29193223

BPG Columns
Operating Instructions
Original instructions
cytiva.com
Table of Contents
Table of Contents
1 Introduction ........................................................................................................ 4
1.1 About this manual ......................................................................................................................................... 5
1.2 Important user information ....................................................................................................................... 6
2 Safety instructions ............................................................................................. 8

2.1 Safety precautions ........................................................................................................................................ 9
2.2 Symbols and abbreviations ........................................................................................................................ 15
2.3 Emergency procedures ............................................................................................................................... 17
3 Column descriptions .......................................................................................... 18

3.1 Overview ............................................................................................................................................................ 19
3.2 BPG 100, 140 and 200 columns ................................................................................................................ 22
3.3 BPG 300 column ............................................................................................................................................. 25
3.4 BPG 450 column ............................................................................................................................................. 28
3.5 Accessories ...................................................................................................................................................... 31
4 Installation .......................................................................................................... 33
4.1 Site requirements .......................................................................................................................................... 34
4.2 Transport .......................................................................................................................................................... 35
4.3 Unpacking ......................................................................................................................................................... 36
4.4 How to level the column .............................................................................................................................. 38
4.5 Grounding kit ................................................................................................................................................... 40
4.6 Setup ................................................................................................................................................................... 42
5 Operation ............................................................................................................ 45
5.1 Preparation ...................................................................................................................................................... 47
5.1.1 Column flow packing method ................................................................................................................. 49
5.1.2 Slurry preparation ........................................................................................................................................ 50
5.2 Column packing .............................................................................................................................................. 53
5.2.1 Preparing the column ................................................................................................................................. 54
5.2.2 Preparing the adapter ................................................................................................................................ 57
5.2.3 Packing the column - first step ............................................................................................................... 62
5.2.4 Packing the column - second step ........................................................................................................ 67
5.2.5 Storage and unpacking of packed columns ...................................................................................... 70
5.2.6 Pressure flow curve ..................................................................................................................................... 71
5.3 Performance evaluation of the column ................................................................................................. 72
6 Maintenance ....................................................................................................... 75
6.1 Service and preventive maintenance .................................................................................................... 77
6.2 Replacements of nets and O-rings .......................................................................................................... 79
6.2.1 Replacements in the adapter .................................................................................................................. 81
6.2.2 Replacements in the column ................................................................................................................... 87
6.3 Replacement sealing washer .................................................................................................................... 98
6.4 Fitting a packing extension tube ............................................................................................................. 99
6.5 Disassemble and assemble the adapter ............................................................................................... 101
6.6 Leakage test .................................................................................................................................................... 113
6.7 Cleaning ............................................................................................................................................................ 115
2 BPG Columns Operating Instructions 29193223 AB

Table of Contents
6.8 Storage .............................................................................................................................................................. 119
7 Troubleshooting ................................................................................................ 121
8 Reference information ....................................................................................... 124

8.1 Specifications .................................................................................................................................................. 125
8.1.1 BPG 100 ............................................................................................................................................................ 127
8.1.2 BPG 140 ............................................................................................................................................................ 128
8.1.3 BPG 200 ............................................................................................................................................................ 129
8.1.4 BPG 300 ............................................................................................................................................................ 130
8.1.5 BPG 450 ............................................................................................................................................................ 131
8.2 Chemical resistance ..................................................................................................................................... 132
8.3 Abbreviations .................................................................................................................................................. 135
8.4 Recycling information .................................................................................................................................. 136
8.5 Regulatory information ............................................................................................................................... 137
8.5.1 Contact information .................................................................................................................................... 138
8.5.2 European Union and European Economic Area .............................................................................. 139
8.6 Health and Safety Declaration Form ...................................................................................................... 140
Index ........................................................................................................................... 142
BPG Columns Operating Instructions 29193223 AB 3

1 Introduction
1 Introduction
About this chapter

This chapter contains important user information, descriptions of safety notices, and
the intended use of BPG columns.
In this chapter
Section See page
1.1 About this manual 5
1.2 Important user information 6

1 Introduction
1.1 About this manual
1.1 About this manual

Purpose of this manual
The Operating Instructions provide you with the information needed to install, operate,
and maintain the product in a safe way.
Scope of this document

This document describes the basic components and principles of operation of BPG
columns. Specific configuration information can be found in the product documenta-
tion provided with each column and on the column label.
Detailed information regarding process systems, media and buffer tanks is not covered
in this document.

1 Introduction
1.2 Important user information

Read this before operating the
product
All users must read the entire Operating Instructions before installing, oper-
ating or maintaining the product.
Always keep the Operating Instructions at hand when operating the product.
Do not install, operate, or perform maintenance on the product in any other way than
described in the user documentation. If you do, you may be exposed or expose others
to hazards that can lead to personal injury and you may cause damage to the equip-
ment.
Intended use of the product

BPG columns are intended for industrial applications that demand high standards of
hygiene. BPG columns can be used in conjunction with BioProcess™ Chromatography
media. The pressure specifications and low flow resistance make these columns suit-
able for use with BioProcess media in ion exchange, hydrophobic interaction, affinity
and gel filtration chromatography.
BPG columns are intended for production use only and should not be used for diag-
nostic purposes in any clinical or in vitro procedures.
Prerequisites
In order to operate the equipment in the way it is intended, the following prerequisites
must be fulfilled:
• You must have read and understood the safety instructions in the user documenta-
tion.
• You must have experience of working with pressure vessels.
• You must have knowledge of the entire system and process that the column is part
of.
• All operations should be performed by qualified personnel who are adequately
trained.
Definitions
This user documentation contains safety notices (WARNING, CAUTION, and NOTICE)
concerning the safe use of the product. See definitions below.

1 Introduction
WARNING
WARNING indicates a hazardous situation which, if not avoided,
could result in death or serious injury. It is important not to proceed
until all stated conditions are met and clearly understood.
CAUTION
CAUTION indicates a hazardous situation which, if not avoided,
could result in minor or moderate injury. It is important not to
proceed until all stated conditions are met and clearly understood.
NOTICE
NOTICE indicates instructions that must be followed to avoid
damage to the product or other equipment.
Notes and tips

Note: A note is used to indicate information that is important for trouble-free and
optimal use of the product.
Tip: A tip contains useful information that can improve or optimize your proce-
dures.

2 Safety instructions
About this chapter

This chapter describes safety precautions and the label attached to the equipment. In
addition, the chapter describes emergency and recovery procedures.
Important
WARNING
Before installing, operating or maintaining the product, all
users must read and understand the entire contents of this
chapter to become aware of the hazards involved.
In this chapter
Section See page
2.1 Safety precautions 9
2.2 Symbols and abbreviations 15
2.3 Emergency procedures 17

2.1 Safety precautions

About this section
The safety precautions in this section are grouped in the following categories:
• General precautions, on page 9
• Flammable liquids and explosive environment, on page 10
• Personal protection, on page 11
• Installing and moving, on page 11
• Operation, on page 12
• Maintenance, on page 13
General precautions
WARNING
Do not operate the product in any other way than described in the
user documentation.
WARNING
Only properly trained personnel may operate and maintain the
product.
WARNING
Do not use any accessories not supplied or recommended by
Cytiva.
WARNING
Pressure may remain in the column and tubing for an extended
period after shutdown. Make sure the system is depressurized
before opening or disconnecting.

Flammable liquids and explosive

environment
WARNING
EXPLOSION HAZARD! If cleaning the column in a potentially
explosive atmosphere, prevent static electric charges building up.
Do not rub excessively using a dry cloth and over large areas at a
time. Instead, clean the column using a damp cloth with slow
movements across small areas at a time.
WARNING
EXPLOSION HAZARD! Some of the chemicals used with BPG
columns may be flammable under certain conditions. Make sure to
use chemicals only under conditions where they are not flam-
mable. Refer to local and/or national classifications of flammable
liquids.
WARNING
EXPLOSION HAZARD! When the BPG column is operated or
maintained in a potentially explosive atmosphere, it MUST be
properly grounded to avoid static discharge.
WARNING
EXPLOSION HAZARD! When connecting the grounding cable,
make sure that there is no explosive atmosphere present. Static
electricity discharge could occur when connecting the grounding
cable.

Personal protection
WARNING
For personal safety during the transportation, installation, opera-
tion, maintenance and service of columns, use protective glasses
and other personal protective equipment appropriate for the
current application at all times. The following personal protective
equipment should always be available:
• Protective glasses
• Working gloves to protect against sharp edges
• Protective footwear, preferably with steel toe-cap
• Disposable gloves
Always use clean disposable gloves when manually handling parts.
WARNING
When using hazardous chemical and biological agents, take all
suitable protective measures, such as wearing protective glasses
and gloves resistant to the substances used. Follow local and/or
national regulations for safe operation and maintenance of the
BPG columns.
Installing and moving
WARNING
Only personnel with appropriate training may move and unpack
the delivery crates. All moving and unpacking must be performed
according to local regulations.
Even if the safety instructions in the user documentation are
followed, it is the customers' responsibility to guarantee the safety
of the personnel working with BPG columns.
WARNING
Move transport crates. Make sure that the lifting equipment has
the capacity to safely lift the crate weight. Make sure that the crate
is properly balanced so that it will not accidentally tip when moved.

WARNING
Have a minimum of two people on hand, as well as the appropriate
lifting equipment for the application, such as a hoist, when moving
or lifting a heavier BPG column, i.e., BPG 300 or 450.
WARNING
Be careful not to unscrew the wheels too far. If they are unscrewed
totally the column may fall and cause injury or damage. Make sure
that the wheel support is fully engaged into the stand before start
leveling.
WARNING
Make sure that the wheel support is fully engaged into the stand
before start moving.
CAUTION
After moving the column to its resting position, lock the wheel
brakes and level the column. Place the column on an even floor
area.
CAUTION
Make sure that the safety equipment's (for example pressure relief
valve or rupture disc) connecting tubing has an unrestricted flow
path and connected to waste.
Operation
WARNING
The working pressure of the column should never exceed its design
pressure, otherwise there is a risk of personal injury and damage to
the column. Always use appropriate safety equipment, for example
pressure relief valve or rupture discs.

WARNING
There should NEVER be air or gas under pressure in the column.
WARNING
Pay extra attention when fitting the adapter to the column as there
is a risk of crushing your hands.
CAUTION
Do not use chemicals with temperatures above the specified limits.
Maintenance
WARNING
Thoroughly check the glass column tube to make sure that it is not
cracked or has been severely scratched. Pay extra attention to the
end parts of the column tube. The pressure specifications are only
valid if the glass tube is undamaged.
WARNING
Decontaminate before maintenance. To avoid personnel being
exposed to potentially hazardous substances, make sure that the
column is properly decontaminated and sanitized before mainte-
nance or service.
CAUTION
For continued protection against injury risks due to fluid jets, burst
pipes or explosive atmosphere, the piping system must be tested
for leakage at maximum pressure when the following situations
apply:
• After assembly or maintenance
• Before operation or Cleaning-In-Place (CIP)

CAUTION
After assembly, check that the column is tight and not leaking.
CAUTION
Removing the top plate and adapter often causes spillages and
splashing. Remove any spillage on the floor immediately to mini-
mize the risk for slipping accidents.

2.2 Symbols and abbreviations

Introduction
This section describes the information on the system label and other safety or regula-
tory labels that are attached to the product.
System label
The system label is located on the back of the equipment. The system label identifies
the equipment and shows electrical data, regulatory compliance, and warning
symbols.
Description of symbols on the system

label
The following symbols and text may be present on the system label:
Label text Description

Code number Code number identifying the column as a BPG column
product.
Serial number Serial number.
Mfg date Year and month the instrument was produced (YYYY/
MM).
Tare mass Weight of the instrument when empty. (Tare mass is not
always presented on the label.)
Operating temp. TS Permitted operating temperature range.
Design pressure PS Pressure the instrument has been designed to tolerate.
PED fluid group / cat. Pressure Equipment Directive fluid group and category
classification.
Warning! Read the user documentation before using the
system.
The equipment complies with applicable European direc-

tives.
Applicable to BPG 200/950, 300/500, 300/750, 300/950,
450/500, 450/750 and 450/1000.

Placement of system label

The system label is placed on the flange on the column.

2.3 Emergency procedures
2.3 Emergency procedures

Precautions
WARNING
Emergency shutdown
The operation of BPG columns is controlled by the equipment that is connected to the
column. All operations are therefore stopped by stopping the operation of the
connected equipment.
If the column is connected to a system, the procedure for the emergency shutdown of
the system should be followed. This may involve pressing the EMERGENCY STOP
button to stop the operation of the equipment controlling the pumping system
connected to the column. Alternatively, the process system may require to be paused
or shutdown. Follow the manufacturer's instructions for the system.
Restart
To restart after an emergency shutdown:
Step Action
1 Make sure that the condition that was the root cause of the emergency stop
has been corrected.
2 If an EMERGENCY STOP button was used to shutdown the equipment

connected to the column, reset this before restart.
3 Reduce pump settings prior to reactivation to avoid immediate high pres-

sure increases.
4 Restart the equipment connected to the column.

3 Column descriptions
About this chapter

This chapter contains a general description of BPG columns and illustrations of the
BPG models.
In this chapter
Section See page
3.1 Overview 19
3.2 BPG 100, 140 and 200 columns 22
3.3 BPG 300 column 25
3.4 BPG 450 column 28
3.5 Accessories 31

3.1 Overview
3.1 Overview
Introduction
The BPG column can be used conjunction with a BioProcess system or as a stand-alone
unit connected to a pump. There are five sizes of the BPG columns.
The BPG columns are listed in the table below with the internal diameters, specific
design pressure and the various heights.
Characteristics BPG 100 BPG 140 BPG 200 BPG 300 BPG 450
Internal diameter 100 140 200 296 446
(mm)
Design pressure 8 6 6 4 2.5
(bar)
Column height 500, 750 , 500, 750 , 500, 750 , 500, 750 , 500, 750 ,
(mm) 950 950 950 950 1000
For detailed specifications of the BPG columns, refer to Column specifications, on page
125.
All BPG columns are equipped with wheels with brakes, except the BPG 100 column, for
which wheel with brakes are an option.

3.1 Overview
Column material
The materials used in the BPG columns are compatible with the solvents most
commonly used in chromatography separations of biomolecules, column mainte-
nance and cleaning processes. All polymeric materials in contact with the process
stream meet the requirements for USP class VI, described in USP <88> Biological Reac-
tivity Tests, In Vivo.
Adapter and rods

The adapter top plate and the adjuster nut are bolted to the column flange. The
adjuster nut is allowed to rotate on a bushing ring in the adapter top plate, altering the
height of the adapter plate in the column tube.
The adapter O-ring forms a seal between the adapter and the glass column when it is
compressed by the sealing unit. This is controlled by the seal adjuster knob.
The rods hold the flange and end-piece to the glass tube which is sealed by O-rings. The
O-rings effectively isolate the glass tube from direct contact with the stainless steel
components.
Nets
The column is delivered with 23 μm (PolyPropylene) nets. For media with an average
particle diameter <70 μm, change to 10 μm (Polyamide), or 12 μm (PEEK) in both
adapters and end-pieces. For big beads use 54 μm nets, see Recommended net sizes,
on page 48.
Connections
BPG columns can be connected to different types of tubing and systems. To protect
the packed bed it is advisable to have isolation valves on both the inlets and outlets of
the column.

3.1 Overview
Explosive atmosphere
Perform a risk assessment for any risks due to the process or process environment.
Evaluate the effects the use of the product and the operational processes may have on
the classification of the hazardous area. The process might cause the area to increase
or the zone classification to change. Implement the risk reduction measures needed,
including use of personal protective equipment.
If potentially explosive liquids will be used with BPG columns consult local authorities
regarding local rules and regulations before installing or operating.
The column can be placed in an area with explosive atmosphere according to EU ATEX
workplace directive, Zone 2.
Definition of the zone A place in which an explosive atmosphere consisting

according to the direc- of a mixture with air of flammable substances in the
tive: Zone 2 form of gas, vapour or mist is not likely to occur in
normal operation but, if it does occur, will persist for a
short period only.
The column does not fall under the ATEX equipment directive, and as a result it is not
marked with EX labels, but it fulfills the explosion safety requirements in the directive’s
harmonized standards, for example EN 13463-1 or EN ISO 80079-36.
This is valid for the BPG column in its standard execution and with a grounding cable,
see Grounding kit, on page 32. Any additional accessory or equipment must carry EX-
marking or be assessed to be in conformity with the EU ATEX workplace directive.

3.2 BPG 100, 140 and 200 columns

BPG parts
The image below shows the parts in BPG column 100, 140 or 200.

Part Description
1 Seal adjuster knob
2 Adapter tube, outer
3 Adapter height adjuster
4 Domed nut
5 Adapter top plate
6 Flange
7 Column glass tube
8 Support rod
9 Adapter plate
10 O-ring adapter
11 Net (adapter)
12 O-ring bottom
13 Net (column)
14 End-piece
15 Stand

Column and adapter
Part Description
1 Column
2 Adapter

3.3 BPG 300 column
3.3 BPG 300 column

BPG parts
The image below shows the parts in BPG 300 column.

3.3 BPG 300 column
Part Description
4 Lifting grip
5 Domed nut
6 Adapter top plate
7 Flange
8 Column glass tube
9 Support rod
10 Adapter plate
11 O-ring adapter
12 Net (adapter)
13 O-ring bottom
14 Net (column)
15 End-piece
16 Stand

3.3 BPG 300 column
Column and adapter
2
1
Part Description
1 Column
2 Adapter

3.4 BPG 450 column
3.4 BPG 450 column

BPG parts
The image below shows the parts in BPG 450 column .
Part Description

3.4 BPG 450 column
Part Description
4 Eyebolt
5 Adapter top plate
6 Flange
7 Column glass tube
8 Support rod
9 Adapter plate
10 O-ring adapter
11 Net (adapter)
12 O-ring bottom
13 Net (column)
14 End-piece
15 Stand

3.4 BPG 450 column
Column and adapter
Part Description
1 Column
2 Adapter

3.5 Accessories
3.5 Accessories
Packing extension tube
Packing extension tubes are available for all BPG columns. The packing tubes are made
of glass for BPG 100 to 300, and of stainless steel for BPG 450. The illustration below
shows two examples of extension tubes.
Pressure relief valve

The pressure relief valve is connected between the pump and column inlet. It releases
pressure if the calibrated value is exceeded. It is to be used if the column may exceed its
design pressure and no other safety equipment is included in the chromatographic
system to prevent this.
Air trap
The air trap kit includes the air trap, bracket, steel valves, clamps and gaskets.
For air traps for BPG 100, 140 and 200, tubing is included. A manually operated valve is
recommended at the top of the air trap as an air outlet control.
Pressure gauge kit

The pressure gauge kit contains a pressure gauge, T-junction, and necessary clamps
and gaskets for sanitary connections.

3.5 Accessories
Grounding kit
To avoid discharge from static electricity, the column can be properly grounded
(earthed). For this purpose, a suitable grounding kit is available as an accessory and a
grounding connection terminal is provided. The equipment must be grounded when it
is to be used in a potentially explosive atmosphere.

4 Installation
4 Installation
About this chapter

This chapter contains information about BPG installation, site requirements,
unpacking, transport and how to set up the BPG columns for use.
In this chapter
Section See page
4.1 Site requirements 34
4.2 Transport 35
4.3 Unpacking 36
4.4 How to level the column 38
4.5 Grounding kit 40
4.6 Setup 42

4 Installation
4.1 Site requirements
4.1 Site requirements

Space and floor load
For space and floor load requirements, refer to the dimensions and weight information
in the product documentation provided with each column. Make sure the floor can
handle the weight of the BPG columns at fully loaded conditions.
In order to allow convenient working conditions for the operator, sufficient space
should be provided on all sides of the column when it is installed at the intended
production location.
Equipment requirements
When transporting and maneuvering BPG columns and adapters the following equip-
ment is recommended:
• Suitable lifting equipment (crane or hoist) for the larger BPG columns, see Moving
the adapter BPG 300 and BPG 450, on page 76.
• Lifting eyebolts.
Ambient environment
The following should be avoided:
• Direct sunlight
• Vibrations
• Corrosive gas
• Dust

4 Installation
4.2 Transport
4.2 Transport
Check the delivery
On receipt of the crate check to see if there is any apparent damage to the crate and
the equipment. When unpacking BPG columns check that all equipment is enclosed in
the transport crate according to the packing list.
If any damage is observed do not continue with the installation of the column, record
this on the receiving documents. Contact your Cytiva representative for advice and
further instruction.
Transport in crate
WARNING
Move transport crates. Make sure that the lifting equipment has
the capacity to safely lift the crate weight. Make sure that the crate
is properly balanced so that it will not accidentally tip when moved.
Use proper lifting device with a minimum capacity to match the empty weight of the
column plus the crate. If maneuvering is performed with a crane or hoist, keep lifting
heights to the minimum necessary to make sure that there is sufficient clearance
space to allow passage of the column when lifted from the floor. Refer to the product
documentation for information about column weight and dimensions for each column.
Moving the column
WARNING
Make sure that the wheel support is fully engaged into the stand
before start moving.
Be careful not to unscrew the wheels too far before moving. If they are unscrewed
totally the column may fall and cause injury or damage. The wheels should be fully
engaged into stand before moving the column.

4 Installation
4.3 Unpacking
4.3 Unpacking
Tools required for crate unpacking
The following tools are recommended when removing larger BPG columns from the
transport crate:
• Suitable lifting equipment, such as a crane or hoist
• Round slings or lifting chains with the capacity to safely lift the weight of the column
• Lifting eyebolts
• Screwdriver
• Suitable wrench or ratchet wrench
Crate unpacking
WARNING
Only personnel with appropriate training may move and unpack
the delivery crates. All moving and unpacking must be performed
according to local regulations.
Even if the safety instructions in the user documentation are
followed, it is the customers' responsibility to guarantee the safety
of the personnel working with BPG columns.
Unpack the BPG 300 and 450 columns by following the unpacking instructions
attached to the outside of the transport crate. In the absence of unpacking instruc-
tions, unpack BPG column from the transport crate as follows:
Step Action
1 Move the crate to a protected indoor location. Make sure that the transport
crate is located on a level and even floor surface.
2 Remove the screws marked with black paint from the side panel of the trans-
port crate, and carefully remove that side panel from the crate.
3 Remove the top panel from the crate.
4 Remove the enclosed accessories that are packed inside the crate.
5 Place the ramp that is provided inside the crate on the floor surface.
6 Secure the ramp tightly against the open side of the crate. Make sure there is
a smooth transition between the bottom of the crate and the floor.
7 Release the column wheel brakes and carefully roll the column out of the
crate and onto the floor.

4 Installation
4.3 Unpacking
Positioning the column
WARNING
The BPG column can be rolled to the intended operational location on its own wheels,
or the larger columns can be lifted into position using a crane or hoist.
Tip: It is recommended to wrap PTFE tape around the threads of the eyebolts
used with BPG column.
Cleaning column surfaces

Before beginning to prepare the column for operation it is recommended that any
surface residues acquired during transport or removal of the column from the packing
crate are removed using warm water and/or 20% ethanol.

4 Installation
4.4 How to level the column

Precautions
WARNING
Be careful not to unscrew the wheels too far. If they are unscrewed
totally the column may fall and cause injury or damage. Make sure
that the wheel support is fully engaged into the stand before start
leveling.
CAUTION
After moving the column to its resting position, lock the wheel
brakes and level the column. Place the column on an even floor
area.
NOTICE
Make sure that the column is located on a level even surface.
Lock the wheels of BPG columns

After positioning the column in its operational location, lock the wheels as illustrated
below.

4 Installation
Use the level adjusters

Before operation the column must be level. Use a spirit level to check if the column is
level. Measure in two positions perpendicular to one another and repeat the measure-
ments at several points around the exterior of the column.
If column is not level screw all wheels fully into the stand and then level on the correct
wheel to avoid adjusting any wheel to far out.
• If the column must be raised, turn the column leg level adjustor clockwise.
• If the column must be lowered, turn the column leg level adjustor counter-clock-
wise.
Check if the column is level after adjusting each leg. Adjust the column legs in turn until
the column is level.

4 Installation
4.5 Grounding kit
4.5 Grounding kit

Precautions
WARNING
EXPLOSION HAZARD! When the BPG column is operated or
maintained in a potentially explosive atmosphere, it MUST be
properly grounded to avoid static discharge.
WARNING
EXPLOSION HAZARD! When connecting the grounding cable,
make sure that there is no explosive atmosphere present. Static
electricity discharge could occur when connecting the grounding
cable.
Ground the column

To ground the column, connect one end of the grounding cable to an earth terminal in
the plant grounding network and the other end to the ground terminal located on the
column stand. The column ground terminal has an M6 thread.

4 Installation
4.5 Grounding kit
Illustration
A grounding kit is available as an accessory for BPG columns and a grounding connec-
tion terminal is provided on the column stand as shown below.
Part Function
1 Grounding terminal/cable on BPG 100, 140, 200 and 450 columns.
2 Grounding terminal on BPG 300 column.

4 Installation
4.6 Setup
4.6 Setup
Precautions
WARNING
CAUTION
Make sure that the safety equipment's (for example pressure relief
valve or rupture disc) connecting tubing has an unrestricted flow
path and connected to waste.
NOTICE
The bottom outlet should be above the top of the column to elimi-
nate any chance of a siphon.
NOTICE
A pressure relief valve or rupture disc shall be connected between
the pump and column inlet.

4 Installation
4.6 Setup
Column setup
The column is fitted with a 4-port 4-way valve on the adapter inlet (B), and a 4-port 2-
way valve on the bottom outlet.
• One port (A) on the top has a pressure gauge followed by tubing connected to a
pump. Another port (C) is directed to the waste for purging the pump and column.
• The bottom valve is equipped with tubing on a port perpendicular to the column
outlet that will be directing flow to waste, or recycling water back to the tank.
3
1 A C
2
B
6
4
Flow Description
1 From pump
2 Pressure gauge
3 4-port, 4-way valve
4 Drain
5 4-port, 2-way valve
6 Drain
7 Return

4 Installation
4.6 Setup
Four-port valve positions
Flow Valve position Description

A to C Mobile phase from pump to waste
A C
A to B A C Mobile phase to column
B
A C
B to C A C Column to waste

5 Operation
5 Operation
About this chapter

BPG columns can be used to perform a number of different chromatographic purifica-
tions, depending on the chromatography media and specific methods selected for use.
This chapter provides general instructions on how to prepare and operate BPG
columns.
In this chapter
Section See page
5.1 Preparation 47
5.2 Column packing 53
5.3 Performance evaluation of the column 72
Precautions
WARNING
product.
WARNING
Thoroughly check the glass column tube to make sure that it is not
cracked or has been severely scratched. Pay extra attention to the
end parts of the column tube. The pressure specifications are only
valid if the glass tube is undamaged.
WARNING
There should NEVER be air or gas under pressure in the column.

5 Operation
WARNING
CAUTION
Do not use chemicals with temperatures above the specified limits.
NOTICE
Make sure that any water is WFI, since particles can block and
damage the nets.
NOTICE
Use a harmless fluid in the beginning of the process. This will make
it possible to detect leakage with minimized consequences and the
risk for potential leakage of hazardous fluids is avoided.

5 Operation
5.1 Preparation
5.1 Preparation
About this section
This section contains information how to prepare the BPG columns before column
packing. Information and support related to optimal column packing can be obtained
from your Cytiva representative.
In this section
Section See page
5.1.1 Column flow packing method 49
5.1.2 Slurry preparation 50
Location
Take care in selecting the position of the column before proceeding. System, tanks,
drains and handling devices for the larger columns all require space and easy, safe
access during all operations of the column. Take care to route the hoses carefully to
avoid tripping risks and always try to keep hose lengths to a minimum.
Arrange connections
NOTICE
Only use tubing and clamps approved for the maximum pressure of
the column and resistant to the chemicals intended to be used.
Make sure the tubing inner diameter is constant throughout your system setup to
minimize unwanted volume and keep a consistent flow and pressure in the process
stream. The valves, tubing and gaskets used should have the same inner diameter as
the column outlets or slightly larger
Materials
The following materials are suitable for the column packing:
• BPG column with suitable net for the media
• Media about 50% slurry in water
• WFI for packing

5 Operation
5.1 Preparation
Recommended net sizes

It is important to select the appropriate adapter and end-piece nets. Nets are available
in 10, 12, 23 and 54 µm porosities. Improperly sized nets can lead to back pressure
problems or media leakage. The table below shows recommended net sizes.
Net size (µm) Material Bead size (µm)

10 PA1 30 to 70
12 PEEK2 30 to 70
23 PP3 90 (FF-media)
54 PP3 120 (BB-media)
1 Polyamide
2 Polyetheretherketone
3 Polypropylene
Packing extension tube

If a packing extension tube is used it will either remain in place for very long bed
heights, or be removed during the packing procedure when the media is below the top
of the column tube.

5 Operation
5.1 Preparation
5.1.1 Column flow packing method
5.1.1 Column flow packing method
Packing flow rate

Packing flow rates should be found in the leaflet supplied with the media. The optimum
packing flow rates is dependent on temperature, type of media,concentration and the
packing method. Consequently, the optimum flow rate or rates must be determined
empirically by producing a pressure/flow rate curve.
It is advisable to do a pressure flow curve with a similar volume media that will be
packed in the column.
Method description
Constant flow packing is a two step method that is applicable to all types of media.
Assuming a suitable liquid delivery system is available, the main requirements are that
the column must have a moveable adapter and a pressure rating which allows the
optimal packing flow rate to be obtained.
1. In the first step, a low flow rate is used to form a loosely packed bed from the slurry.
This allows the adapter to be lowered the majority of the distance towards its final
packed position with little or no expansion of the bed during travel time.
2. In the second step, the flow rate is increased to a predetermined optimal value to
obtain the desired packing density. When flow is stopped, the proximity of the
adapter to the bed minimizes the time necessary to successfully reposition the
adapter to stabilize the packed bed.

5 Operation
5.1 Preparation
5.1.2 Slurry preparation
Slurry concentration
Mix the packing buffer with the media to form a 50% slurry, i.e., sediment volume/total
slurry volume = 0.50. This slurry volume is appropriate for BioProcess media.
Make sure that the correct amount of slurry is used to end up with the target bed
height and optimal bed compression.
More information regarding how to determine slurry concentration using the Slurry
Concentration Kit can be found in Instruction 29112530.
Compression factor
The media needs to be compressed slightly to maintain a stable packed bed. The multi-
plication factor to convert packed bed volume to settled media volume is the compres-
sion factor.
Compression Factor CF = (gravity settled bed height)/(packed bed height)
Compression factor should read for most Cytiva media, between 1.10 and 1.15.
Amount of media and slurry volume

The amount of media required for packing the column can be calculated using the
following formula:
A = (π × r2 × H × Cf/1000)
Where:
A = volume of media needed, liter
π = 3.14
r = radius of column (half the diameter), cm
H = bed height in column, cm
Cf = Compression factor of media
The 50% slurry volume is calculated as: A/0.5
Media quantities required per cm bed

height
The table shows the amount of media per centimeter of packed bed. The compression
factor is 1.15.
BPG column Column diameter (π × r2 × H × Cf/1000) Amount of media1 per cm of

(mm) packed bed
BPG 100 100 78.5 × 1.15 90.3 ml
BPG 140 140 154 × 1.15 177 ml

5 Operation
5.1 Preparation
BPG column Column diameter (π × r2 × H × Cf/1000) Amount of media1 per cm of

(mm) packed bed
BPG 200 200 314 × 1.15 361 ml
BPG 300 296 0.707 x 1.15 812 ml
BPG 450 446 1.59 x 1.15 1.83 l
1 DEAE Sepharose™ FF, Q Sepharose FF, CM Sepharose FF, S Sepharose FF, Phenyl Sepharose FF, Sepharose 4 FF and
6 FF, Sephacryl™ High Resolution, Superdex™ 75 and 200 prep grade, Q and SP Sepharose HP, Phenyl Sepharose HP
Sephadex G-25
The Sephadex™ G25 media swell 4 to 5 times (1 g = 4-5 ml of media). The table shows
the amount of media per centimeter of packed bed.
Note: The figures in the table below are based on four-fold swelling and that there
can be batch-to-batch variation.
Media Amount of Pack Amount of media per cm of packed bed (g)

media per factor
liter of (PF) BPG BPG BPG BPG BPG
packed 100 140 200 300 450
(l)
column
(g)
Sephadex G-25 250 1.15 l 22.6 43.9 90.3 202.4 453
C and M grades
Sephadex G-25 280 1.15 l 22.6 43.9 90.3 202.4 453
grade SF

5 Operation
5.1 Preparation
Prepare the slurry

Follow the instructions below to mix the slurry for column packing:
Step Action
1 Make sure that the slurry is homogeneous.
2 Pour the slurry into a suitable container for pouring into the column.
3 Check the slurry concentration.

5 Operation
5.2 Column packing
5.2 Column packing

About this section
This sections describes the steps that must be taken in order to pack the BPG columns.
In this section
Section See page
5.2.1 Preparing the column 54
5.2.2 Preparing the adapter 57
5.2.3 Packing the column - first step 62
5.2.4 Packing the column - second step 67
5.2.5 Storage and unpacking of packed columns 70
5.2.6 Pressure flow curve 71

5 Operation
5.2 Column packing
5.2.1 Preparing the column
Column preparation
Operation of the adapter, integrity of the adapter O-ring and porosity of the support
nets should be checked before starting. The column tube and all internal surfaces must
be clean.
Remove air from bottom net

Every effort must be made to remove air from bottom net and tubing. Air is removed
from under the bottom net using suction and about 1 cm buffer left in the column.
Step Action
1 Make sure that the column is level, see Section 4.4 How to level the column,
on page 38.
2 Make sure that the column is grounded, see Section 4.5 Grounding kit, on
page 40.
3 Wet the column net with 20% ethanol and pour 2 to 3 cm of packing buffer
into the column with the bottom valve closed and the outlet to drain.

5 Operation
5.2 Column packing
Step Action
4 Open the outlet valve at the bottom of the column and allow liquid and air to
drain from the bottom distributor plate.
5 Close the bottom valve.
6 Connect a pipe to a pump, or a vacuum line, and lower the pipe down onto
the surface of the bottom net.
7 Start the suction to remove any remaining air from under or on the surface
of the bottom net.

5 Operation
5.2 Column packing
Step Action
8 Continue until there is only 1 cm of buffer in the column.
Result:
The column tube is now ready for packing.

5 Operation
5.2 Column packing
5.2.2 Preparing the adapter
Position and seal the adapter

Step Action
1 Connect the 4-port, 4-way valve, to the top of the adapter.
2 Position the valve from pump to waste (A to C) to purge air and start the
pump.
A C
3 Stop the pump once all air is removed.
4 Position the valve from adapter to waste (B to C).
A C

5 Operation
5.2 Column packing
Step Action
5 Carefully pour the homogenous slurry into the column. Make sure that no air
bubbles are trapped within the slurry and the bottom outlet is higher than
the column.
6 Measure the height (H) from the top of the column to the surface of the
slurry.
H cm

5 Operation
5.2 Column packing
Step Action
7 Set the adapter height to [H - (0.5 to 1.0)] cm.
A C
H - (0.5 - 1.0) cm
8 Insert the adapter in the column just above the slurry level in the column
with the seal fully relaxed.
A C
H - (0.5 - 1.0) cm
WARNING
Pay extra attention when fitting the adapter to the
column as there is a risk of crushing your hands.

5 Operation
5.2 Column packing
Step Action
9 Secure the column adapter in place.
10 Seal the adapter O-ring by turning the seal adjuster knob or handle on the
top of the adapter, clockwise.
11 Wait until a small clear zone is visible above the slurry.

5 Operation
5.2 Column packing
Step Action
12 Move the height adjuster handle clockwise to position the adapter

0.1 to 0.5 cm below the top surface of the liquid.
A C
13 Result:
Air, and then liquid, is pushed up through the adapter and out to waste as the
adapter is pushed downwards.
A C

5 Operation
5.2 Column packing
5.2.3 Packing the column - first step
Description of the first step

The first step of constant flow packing is to form a consolidated bed at a low flow rate
(20 – 30 cm) and position the adapter near the final bed height. The top valve is placed
in position to allow buffer to exit from the column up through the adapter to waste. The
adapter is again lowered until air under the net and in the adapter shaft has been
purged. The pump is started and the tubing to the column also purged of air without
changing the valve position.
The flow should be quickly adjusted to approximately that desired for the first step and
then the top valve turned to place the column inline with the pump. The bottom valve is
immediately opened to waste or recycle, and the flow rate adjusted as needed. After
the bed height of the media stabilizes in one to two column volumes (1 to 2 CVs). The
adapter O-ring is loosened so buffer will pass above it, and the adapter lowered to
0.5-1.0 cm above the top of the bed.
The O-ring is retightened and the adapter lowered to 0.1 to 0.2 cm above the bed with
buffer passing out the top valve to waste. The top valve should be turned to place the
pump in line with the column again and the bottom valve opened. The user should
proceed immediately to the second packing step.
First step
Step Action
1 Remove any air in the hoses by positioning the valve from system to waste (A
to C) and start the pump.
A C

5 Operation
5.2 Column packing
Step Action
2 Position the valve to flow from the pump to the column (A to B) through the
inlet on the adapter, and at the same time open the bottom valve to waste.
A C
3 Check the packing flow rate and adjust as necessary.
4 Stop the pump when the bed has settled, normally one to two column
volumes.
A C

5 Operation
5.2 Column packing
Step Action
5 Close the bottom valve when the pressure in the column has returned to
zero and set the top valve to waste (B to C).
A C
6 Loosen the adapter O-ring seal by turning the adjuster knob, or the seal
adjuster handle, on the top of the column counter-clockwise.
A C

5 Operation
5.2 Column packing
Step Action
7 Lower the adapter quickly by turning the height adjuster handle clockwise.
The buffer between the sedimented bed and the adapter passes between
the adapter O-ring and the column wall. During this operation the bed will
start to rise. Stop lowering the adapter when it is 0.5 to 1.0 cm above the bed.
A C
0.5 - 1.0 cm
8 Seal the adapter O-ring by turning the knob, or seal adjuster handle, clock-
wise and lower the adapter to 0.1 or 0.2 cm above the bed.
A C
0.1 - 0.2 cm

5 Operation
5.2 Column packing
Step Action
9 Result:
The buffer is passing out the top valve to waste (B to C).
A C
10 Close the top valve.

5 Operation
5.2 Column packing
5.2.4 Packing the column - second step
Description of the second step

The second step of flow packing is to compress the bed using a flow rate predeter-
mined from the pressure flow curve data, and rapidly set the adapter position to main-
tain the packed bed density. The pump is adjusted quickly to the desired flow rate to
obtain the final bed height, and the integrity of the adapter O-ring seal checked and
further tightened if necessary. When the bed has stabilized in about 1 CV, the bed
height is marked on the column tube. The top valve is turned to place the pump out of
line with the column at the same time as the bottom column valve is closed.
The adapter must be rapidly lowered into place with minimal disturbance of the packed
bed. This should be done with the adapter O-ring just loose enough that buffer exits out
of the top valve and not around the O-ring. Once the adapter is in place, the O-ring
should be retightened immediately. If the packed bed decompresses significantly
above the marked bed height before the adapter can be lowered in place, this second
packing step can be repeated once. Wait at least 5 minutes for the bed to cure before
moving the column.
Second step
Step Action
1 Open the bottom valve and put the top valve from pump to column (A to B)
and start the pump. This will move the bed down and once stabilized the
level should be marked on the tube.
A C
2 Stop the pump.

5 Operation
5.2 Column packing
Step Action
3 When the pressure reaches zero, position the top valve to waste (B to C), and
move the adapter down, pushing into the media until the adapter reaches
the mark on the tube.
A C
4 Close the top valve and the bottom valve.
5 Result:
The column is now packed.
6 Test the packed column, see Section 5.3 Performance evaluation of the
column, on page 72.

5 Operation
5.2 Column packing
Remove buffer from adapter

The design of BPG columns allows buffer to be on top of the adapter while packing.
When packing is completed and the O-ring is sealed it is essential that all buffer (espe-
cially salt) is removed from that space to avoid corrosion over time. For more informa-
tion about NaCl, see Section 8.2 Chemical resistance, on page 132.
Follow the instructions below to remove buffer from adapter:
Step Action
1 Siphon or pump out the buffer.
A C
2 Wash, if necessary.
3 Repeat.

5 Operation
5.2 Column packing
5.2.5 Storage and unpacking of packed columns
5.2.5 Storage and unpacking of packed columns
Storage
Both outlets of the column should be sealed after equilibrating the column in storage
buffer. The column must be stored in a temperature-controlled environment with
water or 20% ethanol above the adapter O-ring to prevent it from drying out and
cracking. Covering the column with a clean plastic bag helps prevent dust and debris
from accumulating on exterior surfaces.
Unpacking
Place several centimeters of buffer or water above the adapter O-ring of the BPG
column and fully loosen the wing nut which seals the adapter O-ring. Make sure the
adapter outlet is open to waste. If the adapter is not easily raised, try slightly lowering it
to break the O-ring seal on the glass tube. If the O-ring rolls, tighten the wing nut a bit
to prevent this during adapter ascent. A firm twist on the threaded adapter shaft is also
useful when attempting to unseal the O-ring.
Raise and remove the adapter. If the packed bed is difficult to reslurry, pumping buffer
up through the bottom valve will help to lift and disperse the media more rapidly and
clear the bottom net.

5 Operation
5.2 Column packing
5.2.6 Pressure flow curve
5.2.6 Pressure flow curve
Plot the result

Plot the pressure in the flow rate, as indicated in the below example. The optimal
packing flow rate is about 70% of the maximum flow rate. For some very rigid media
there will be no plateau, in those cases use the flow rate at maximum allowable pres-
sure for the column. The figure below shows the pressure flow curve.
70% 100%
b c
The description of the axis are:

x = Pressure
y = Flow rate
The characters in the table are described below:
Value Meaning
a Packing flow rate
b Packing pressure
c Max pressure

5 Operation
5.3 Performance evaluation of the column

About this section
This section describes how to measure HETP and As. To perform a test of the packed
column, read the information supplied with the media giving you all relevant informa-
tion you need.
Column efficiency test

The efficiency of a column depends on how well it is packed. A poorly packed column
gives rise to uneven flow, resulting in zone broadening and reduced resolution. A
column efficiency test should be performed directly after packing and at regular inter-
vals during the working life of the packed bed. The test is also useful when the separa-
tion performance is seen to deteriorate. Poor test results indicate that the media
should be repacked or replaced.
A widely used method for expressing the efficiency of a packed bed is in terms of the
height equivalent to a theoretical plate (HETP) and the peak asymmetry factor (As).
These values can easily be determined by applying a sample of for example acetone or
NaCl solution to the column.
Measuring HETP and As

Measuring the HETP and As values is the best way to judge the condition of the packed
column. A packed column can look good, but still need repacking for optimal perform-
ance. Always check the column after packing and regularly between runs to ensure
best column performance. Follow the specific instructions for the packed media to
optimize the testing protocol.

5 Operation
Test chromatogram
The following test chromatogram shows the parameters used to calculate peak broad-
ening and peak symmetry.
Ve
y
WVhh
50%
a b
10%
x
The description of the axis are:

x = Volume or time
y = UV absorption or conductivity
HETP calculation
Calculate the height equivalent of a theoretical plate (HETP) from the UV curve or
conductivity curve if sodium chloride is used as sample, as follows:
HETP = L/N
N=5.54 (Ve/Wh)2
Variable Meaning
L Bed height
N Number of theoretical plates
Ve Elution volume
Wh Peak width at half peak height
Ve and Wh are in the same units

5 Operation
Asymmetry calculation
Calculate the asymmetry factor (AS) from the UV curve or conductivity curve if sodium
chloride is used as sample, as follows:
The peak should be symmetrical and the asymmetry factor as close to 1 as possible
(values between 0.8 and 1.8 are usually acceptable). A change in the shape of the peak
is usually the first indication of bed deterioration due to use.
The asymmetry factor As describes the deviation from an ideal Gaussian peak shape
and is calculated from the peak width at 10% of peak height.
As = b/a

6 Maintenance
6 Maintenance
About this chapter

This chapter provides a general overview of cleaning, sanitization, replacements, and
maintenance procedures.
In this chapter
Section See page
6.1 Service and preventive maintenance 77
6.2 Replacements of nets and O-rings 79
6.3 Replacement sealing washer 98
6.4 Fitting a packing extension tube 99
6.5 Disassemble and assemble the adapter 101
6.6 Leakage test 113
6.7 Cleaning 115
6.8 Storage 119
Precautions
WARNING
WARNING
Decontaminate before maintenance. To avoid personnel being
exposed to potentially hazardous substances, make sure that the
column is properly decontaminated and sanitized before mainte-
nance or service.

6 Maintenance
Moving the adapter BPG 300 and BPG

450
WARNING
The adapters on BPG 300 and 450 columns are heavy. For safety reasons, it is recom-
mended that the adapter is lifted with an adapter handling unit or ceiling hoist
connected to the eyebolts on the adapter top plate.

6 Maintenance
6.1 Service and preventive maintenance

Precautions
WARNING
product.
WARNING
Do not use any accessories not supplied or recommended by
Cytiva.
NOTICE
If the column tube or the rods are replaced, the column assembly
must be checked in accordance with local authority instructions.
Service frequency
Regular service and maintenance of the column is necessary to maintain the column in
optimal condition and extend the operational lifetime of column components. Nets
and and worn O-ring seals should be checked regularly for wear.
The requirements for frequency of service and preventive maintenance depend on the
frequency of use of the column and the specific applications performed.
A general recommendation is one service and preventive maintenance visit every two
years, but if columns are in continuous operation an annual visit is recommended.
Contact your local Cytiva representative for information about frequency of service
requirements to suit individual application needs.
Note: Note that the recommendations may not apply to your specific use of the
system. The system owner is solely responsible for establishing applicable
routines for periodic maintenance.
Cleaning before planned

maintenance/service
To ensure the protection and safety of service personnel, all equipment and work areas
must be clean and free of any hazardous contaminants before a Service Engineer
starts maintenance work.

6 Maintenance
Please complete the checklist in the On Site Service Health and Safety Declaration
Form or the Health and Safety Declaration Form for Product Return or Servicing,
depending on whether the instrument is going to be serviced on site or returned for
service, respectively.
Health and safety declaration forms

Health and safety declaration forms are available for copying or printing in the Refer-
ence information chapter of this manual, or on digital media supplied with the user
documentation.
Passivation procedure
It is recommended to include a passivation procedure into a service and preventive
maintenance program. Contact your Cytiva representative for additional information.

6 Maintenance
6.2 Replacements of nets and O-rings

In this section
Section See page
6.2.1 Replacements in the adapter 81
6.2.2 Replacements in the column 87
Nets
Blockage of the nets causes malfunction of the columns and an over-used net can
affect distribution. To prevent blockage follow the instructions listed below.
• When columns are to be re-packed, always check the nets and wash them in a
detergent solution or replace them.
• It is important to select the appropriate adapter and end-piece nets, see Recom-
mended net sizes, on page 48.
• Check the HETP and As regularly to prevent poor performance due to old nets.
O-rings and seals

The sealing O-rings may lose their flexibility with time and need to be replaced regu-
larly. Worn O-rings may not seal properly.
Tip: Cytiva recommends that column O-rings and seals are changed at least
once each year. It may be necessary to change O-rings and seals exposed to
some form of movement in the column more frequently, for example the O-
rings on the adapter.
Liquids required for changing nets

and O-rings
• 100% ethanol for cleaning
• 20% ethanol for lubricating seals
• Warm water (approximately 50°C)

6 Maintenance
Tools required for changing nets and

O-rings
The following tools are recommended when replacing nets and O-rings on the BPG
columns.
Illustration Part
Hex keys
Spanners
Torque wrenches for hexagonal socket fittings and

torque settings of 4, 5, 6, 6.5, 8 Nm
12-point opening sockets
Screwdrivers

6 Maintenance
6.2.1 Replacements in the adapter
Adapter items
Order Part Description BPG 100 - BPG BPG

Top- 140 - 200 300 450
Down
1 Adapter plate Yes Yes Yes
2 O-ring Yes Yes Yes
3 Distribution N/A Yes Yes

plate
4 Snap plug N/A No Yes
5 Support net Yes Yes Yes
6 Net Yes Yes Yes

6 Maintenance
Remove the adapter

The instructions below show an example how to remove the adapter from the column
tube.
Note: The instructions show a BPG 300 column but is applicable even to other BPG
columns.
Step Action
1 Turn the handles on the adjusting handle (or knob) several turns to loosen
the adapter O-ring seal, counter-clockwise.
Tip:
Move the adapter down a few millimeters to loosen the O-ring from the glass
wall. Then move it up as far as it will go.
NOTICE
If the adapter O-ring seal is dry, it may be damaged by
forced movement of the adapter. Wet the O-ring with
water or 20% ethanol solution. Do NOT grease the O-
ring.
2 Remove the domed nuts and washers from the adapter top plate with help of
a spanner. Loosen the screws cross-wise according to the illustration below.

6 Maintenance
Step Action
6
7
3
1
2
4
8
NOTICE
Do not loosen each nut completely in one movement.
Three to four turns per nut is enough to loosen and
remove the nuts.
3
CAUTION
Removing the top plate and adapter often causes spill-
ages and splashing. Remove any spillage on the floor
immediately to minimize the risk for slipping accidents.
Loosen the adapter top plate and lift out the adapter. Remove the adapter
without letting the stainless steel touch the glass.

6 Maintenance
Step Action

6 Maintenance
Replace net in the adapter

The instructions below show an example how to replace the adapter net.
Note: The instructions show a BPG 300 column but is applicable to other BPG
columns.
Step Action
1 Locate the small slot at the base of the net ring.
2 Remove the net by bending it carefully.
Note:
BPG 450: Remove the net by twisting a spatula or similar stainless blunt
blade in the slot.
3 Remove the support net. Be careful not to scratch the adapter as this may
cause leakage.
4 Fit new net items according the order in the table Adapter items, on page 81.

6 Maintenance
Step Action
5 Wet the net with 20% ethanol to remove all air bubbles trapped in the net. If
the net is tight, warm it in 50°C water.
NOTICE
Wet the net with 20% ethanol to eliminate trapped air.
Replace the O-ring in the adapter
NOTICE
Make sure not to scratch any steel surfaces of the column when
removing seals and O-rings. If scratched, the sealing of the column
may be compromised and leakage may result.
Follow the instructions below to replace the O-ring in the adapter.
Step Action
1 Pull the O-ring off.
2 Attach a new O-ring lubricated with 20% ethanol.

6 Maintenance
6.2.2 Replacements in the column
End-piece items
Order Part Description BPG 100 BPG BPG

Top- - 140 - 300 450
Down 200
1 O-ring Yes Yes N/A
2 Guide ring Yes N/A N/A
3 U-shaped seal N/A N/A Yes

on net
4 Net Yes Yes Yes
5 Support net Yes Yes Yes
6 Snap plug N/A N/A Yes
7 Distribution N/A Yes Yes

plate
8 End-piece Yes Yes Yes

6 Maintenance
Torque wrench
Use the torque wrench listed in the following table when securing the column tube to
the end-piece. The torque apply for both ends of rods for BPG 450. For BPG 100 to 300
the rod is threaded into the bottom flange and the same torque apply as for the top.
Tighten the bolts in crosswise sequence according to figures below.
NOTICE
Tighten the nuts and bolts close to the given torque. But do not
exceed the given torque, this could damage the column. Always
use a torque wrench.
BPG Description 100 140 200 300 450
1 Support rod 4 5 5 N/A N/A

3
nuts (4) Nm Nm Nm
4
2
Support rod N/A N/A N/A 6 N/A

6 3
nuts (6) Nm
1 2
4 5
Adapter top N/A N/A N/A 6.5 N/A

6 plate bolts Nm
7
3
1
2
4
8

6 Maintenance
BPG Description 100 140 200 300 450

Support rod 4 5 5 6 N/A
1 3
nuts (4) Nm Nm Nm Nm
4 2
6
End-piece N/A N/A N/A N/A 8
7
domed nuts Nm
1 3
4 2
5 8
6 Support rod N/A N/A N/A N/A 4

7
1 3 bolts top and Nm
bottom.
4 2
5 8

6 Maintenance
Disassemble the column

The instructions below show an example of how to disassemble the column.
columns.
Step Action
1 Loosen the nuts on the flange cross-wise in two steps to avoid damage to
the column tube.
1 3
4
2
2 Remove the flange carefully.
NOTICE
When the flange is removed, the column tube is unsup-
ported. Make sure that the column tube does not tip
over.
3 Use a spanner to unscrew the support rods from the end-piece.

6 Maintenance
Step Action
4 Lift off the glass tube. Carefully place the glass tube on a secure surface.
5 Remove the bolts and washers securing the end-piece to the stand using a
spanner.

6 Maintenance
Step Action
6 Remove the nets and O-ring.

Tip:
To be able to remove the O-rings without damaging other parts, spray 20%
ethanol over the flange or seal, or if that does not work, pour 50°C hot water
into the column and let stand for 15 to 30 minutes. If solvents are not
compatible with EPDM seals, change to seals in FEP/PFR.
NOTICE
Make sure not to scratch any steel surfaces of the
column when removing seals and O-rings. If scratched,
the sealing of the column may be compromised and
leakage may result.

6 Maintenance
Replace nets in the end-piece

The instructions describe how to replace nets in the end-piece.
columns.
Step Action
1 Fit new end-piece items according the order in table End-piece items, on
page 87.
Note:
Wet the net with 20% ethanol to eliminate trapped air.
Note:
BPG 450: Carefully place the column tube on the U-shaped seal. Make sure
the flange is centered on the U-shaped seal.
2 Secure the end-piece to the stand using the bolts and washers.

6 Maintenance
Change top and bottom flange O-

rings in the column
Follow the instructions below to change the top and bottom flange O-rings in the
column.
Step Action
1 Disassemble the column according to Disassemble the column, on page 90.
2 Position and fit the bottom flange O-ring.
3 Put back the glass tube.
4 Position and fit the new top O-ring.
5 Assemble the column according to Assemble the column, on page 95.

6 Maintenance
Assemble the column

The instructions below is an example how to assemble the column.
Note: The instructions shows a BPG 140 column but is applicable to other BPG
columns.
Step Action
1 Carefully place the glass tube on the O-ring in the end-piece. Make sure that
the glass tube does not touch the stainless steel. Align the glass tube with
the bottom net.
Note:
BPG 450: Carefully place the column tube on the U-shaped seal. Make sure
the flange is centered on the U-shaped seal.
NOTICE
The glass tube can crack if touching any stainless steel.
2 Screw the support rods into the end-piece, see Torque wrench, on page 88.
3 Fit the column tube to the end-piece with the bolts.
4 Position the flange so that the support rods fit into the appropriate holes.

6 Maintenance
Step Action
5 Make sure that the O-ring on the flange is aligned with the glass tube.
NOTICE
Twisted or deformed O-rings will cause leakage.
6 Align the inner surface of the glass tube with the inner surface of the flange.
7 Fit the washers and the nuts on the support rods and fingertighten the nuts.
8 Set the torque wrench according to specifications, see Torque wrench, on

page 88.
NOTICE
Do not exceed the given torque, this could damage the
column.
9 Tighten the nuts in cross-wise sequence evenly as shown in Torque wrench,

on page 88.
CAUTION
After assembly, check that the column is tight and not
leaking.

6 Maintenance
Step Action
NOTICE
Failure to use cross and stepwise tightening can damage
the column tube.

6 Maintenance
6.3 Replacement sealing washer
6.3 Replacement sealing washer

Replace the PTFE sealing washer in
valves
Follow the instructions below to replace the sealing washer.
Step Action
1 Unscrew the retaining nut at the bottom of the valve with help of the handle.
2 Insert the new PTFE sealing washer in the positioning disc.
3 Insert the positioning disc in the square of the tapered plug (the PTFE sealing
washer has to touch the body).
4 Insert the dished washer (the larger diameter has to touch the positioning
disc).
5 Tighten the retaining nut until the dished washer is flat.
6 Tighten the retaining nut up 1/4 turn.
7 Stabilize the valve for 24 hours, minimum (as PTFE is a floating material).
8 Screw the retaining nut off 3/8 turn (the tightening torque of the retaining
nut is 1.5 Nm).

6 Maintenance
6.4 Fitting a packing extension tube

Precautions
NOTICE
Do not pack the column to bed heights higher than the length of
the column, i.e., the adapter should seal against the column wall,
NOT against the extension tube wall. A tube length of 10 cm is
required when installing the adapter onto the column tube if the
packing extension is removed.
BPG 100, 140 and 200 columns

The extension tube is assembled when delivered.
Follow the instructions below to fit an extension tube onto the BPG column.
Step Action
1 Remove the adapter from the column and place the extension tube onto the
upper flange. Note that the column rods fit into the open holes in the flange
on the extension tube.
2 Make sure that the thin O-ring is positioned properly in the flange.
3 Use the washers and domed nuts from the column lid to secure the exten-
sion tube to the column.
4 The extended column is now ready for the first packing step in two-step
packing procedures.

6 Maintenance
BPG 300 and 450 columns

The extension tube is assembled when delivered.
Follow the instructions below to fit an extension tube on the BPG 300 column.
Step Action
1 Remove the adapter from the column and place the extension tube onto the
upper flange.
2 Make sure that the thin O-ring is properly positioned in the flange.
3 Use the bolts delivered with the extension tube to secure the extension tube
to the column.
4 The extended column is now ready for the first packing step in two-step
packing procedures.

6 Maintenance
6.5 Disassemble and assemble the adapter

Autoclaving
If the adapter is going to be autoclaved it must be disassembled in its parts. This
section shows how to disassemble and assemble the adapter.
Disassemble the adapter

The instructions below show an example how to disassemble the adapter.
columns.
Step Action
1 Remove the adapter from the column tube according to instructions in

Remove the adapter, on page 82.
2 Remove the adapter screw bolts.
3 Lift off the adapter top plate and adapter screws. Take care not to bend the
inlet tube.
4 The top plate assembly can be disassembled by removing the screws.

6 Maintenance
Step Action
5 Unscrew and remove the cap screws that secure the stopper on the adapter
top plate with the hex key.
6 Remove the stopper.

6 Maintenance
Step Action
7 Remove the stop screw and locking nut from the top plate.
8 Unscrew the bolts on the upper ring of the sealing unit. This will release the
sealing unit and the inner adapter tube.

6 Maintenance
Step Action
9 Carefully unscrew and remove the adjusting knob. This releases the tension
on the spring situated under the inner adapter tube.
10 Remove the top-plate assembly by turning it in a clockwise direction.

6 Maintenance
Step Action
11 Remove the sealing unit and the adapter O-ring.

6 Maintenance
Assemble the adapter

The instructions below is an example how to assemble the adapter.
Note: The instructions show how to assemble a BPG 300 column but is applicable
to other BPG columns.
Step Action
1 Position the bushing ring(s) in the recess on the adapter top plate.
Note:
This image shows BPG 140 column but is applicable to other BPG columns.
2 Using the screws secure the adjuster nut insert to adjuster nut body through
the adapter top plate so that the adjuster nut body turns on the bushing ring.
Note:
This image shows BPG 140 column but is applicable to other BPG columns.
3 Slide the spring (c), inner adapter tube (b) and outer adapter tube (a) onto the
adapter plate.

6 Maintenance
Step Action
4 Fasten the bolts by hand to assemble the outer adapter tube to the adapter
plate.
5 Tighten the bolts with a ring spanner to complete the assembly.

6 Maintenance
Step Action
Torque:
BPG 100 4Nm
BPG 200 4Nm
BPG 300 8Nm
BPG 450 8Nm
6 Slide the sealing unit onto the adapter tube.
7 In a counter clockwise direction, screw down the top plate assembly 5 cm

along the outer adapter tube.

6 Maintenance
Step Action
8 Turn the adjusting knob clockwise onto the top of the adapter until the holes
in the inner adjusting tube are level with the holes in the outer adapter tube.
(See Chapter 3 Column descriptions, on page 18 for respective size of the
adjusting knob.)
9 Use a flashlight and a screwdriver through one hole to lock the inner tube in
position. Then two of the adapter head screws can fasten the sealing unit in
place.

6 Maintenance
Step Action
10 Level the hole in the outer adapter tube with the inner adapter tube with the
help of a fine screwdriver.
11 Remove the screwdriver and fit the third screw.

6 Maintenance
Step Action
12 Tighten the screws with a spanner.
13 Locate the stop screw and locking nut in the top plate.

6 Maintenance
Step Action
14 Turn the stop screw until it engages the groove in the outer adaptor tube,
preventing sideways movement of the tube without preventing vertical
movement.
15 Lock the stop screw in position with the locking nut.
16 Attach the nets on the adapter plate.

6 Maintenance
6.6 Leakage test
6.6 Leakage test

Precautions
CAUTION
For continued protection against injury risks due to fluid jets, burst
pipes or explosive atmosphere, the piping system must be tested
for leakage at maximum pressure when the following situations
apply:
• After assembly or maintenance
• Before operation or Cleaning-In-Place (CIP)
Perform leakage test

Follow the instructions below to test for leakage across the adapter.
Step Action
1 Fill the column with water or sanitizing solution.
2 Position the adapter in the column to the same height as you intend to run
the column.
NOTICE
If the adapter O-ring seal is dry, it may be damaged by
forced movement of the adapter. Wet the O-ring with
water or 20% ethanol solution. Do NOT grease the O-
ring.
3 Seal the adapter with the seal adjuster.
4 Remove the air in the column.

6 Maintenance
6.6 Leakage test
Step Action
5 Close the column outlet valve.

A C
Pressure relief valve
6 Raise the pressure to maximum working pressure. Note that pressure spikes
may cause poor packing (cracking).
A pressure relief valve should be placed between the pump and column.
WARNING
The working pressure of the column should never
exceed its design pressure, otherwise there is a risk of
personal injury and damage to the column. Always use
appropriate safety equipment, for example pressure
relief valve or rupture discs.
7 Wait about 15 minutes until the pressure has stabilized.
8 Check the pressure drop on the pressure gauge. The pressure drop should
not exceed 0.1 bar during 5 minutes. A recorder linked to a pressure
controller gives more precise figures.

6 Maintenance
6.7 Cleaning
6.7 Cleaning
Regular cleaning
Regular cleaning of BPG columns is essential for reliable results. It is important that the
column is kept free from contamination such as microbiological growth and adsorbed
proteins.
Precautions
WARNING
EXPLOSION HAZARD! If cleaning the column in a potentially
explosive atmosphere, prevent static electric charges building up.
Do not rub excessively using a dry cloth and over large areas at a
time. Instead, clean the column using a damp cloth with slow
movements across small areas at a time.
NOTICE
Cleaning. Keep the exterior of the instrument dry and clean. Wipe
regularly with a soft damp tissue and, if necessary, a mild cleaning
agent. Let the instrument dry completely before use.
NOTICE
Make sure that any water is WFI, since particles can block and
damage the nets.
NOTICE
Do not leave O-rings or seals in hot water for longer than the time
period recommended in Cytiva product documentation.
Cleaning agents
All column parts can be cleaned with the most commonly used agents, such as deter-
gents, ethanol, weak acids, sodium hydroxide and high salt concentrations. For special
cleaning agents, please refer to Section 8.2 Chemical resistance, on page 132.
In time, even electropolished 316L stainless steel may show signs of corrosion, espe-
cially if it is in frequent contact with high salt concentrations. Should this occur, repas-
sivation of steel components is recommended and should correct the problem.

6 Maintenance
6.7 Cleaning
Cleaning-in-place (CIP)
One of the most important aspects of production chromatography is the maintenance
of the packed column by cleaning-in-place (CIP) procedures. Regular CIP of the packed
column between production batches is needed to ensure the proper product quality
and the expected media and equipment life. CIP removes precipitated material,
strongly bound substances, and other contaminants from the column bed without
dismantling the column.
Recommended CIP procedure

CIP protocols should be included as part of the general process. CIP of the media
should take place when required, for example after each production batch, when an
unusually high back-pressure is generated over the column, or when a change in peak
symmetry or discoloration of the media is observed. See the recommended cleaning
procedures in the table below.
Media Contaminants Suggested procedure Follow-up

Sepharose Soluble proteins/ Wash with 0.5 column volumes of Regenerate to start
Fast Flow and electrostatic filtered 2 M NaCl for a contact time conditions. See
Sepharose interaction. 10 to 15 minutes. instructions for
High Perform- respective media.
ance based
media Soluble proteins/ Wash with 2 to 3 column volumes of Regenerate to start
hydrophobic filtered low ionic strength buffer or conditions. See
interaction. water for a contact time of 10 to 15 instructions for
minutes, or respective media.
wash with 1 column volume of 30%
isopropanol for a contact time of
10 to 15 minutes, or
wash with 2 to 4 column volumes of
0.5 to 1 M NaOH for a contact time of
1 to 2 hours.
Precipitated Wash with 2 to 4 column volumes of 1 If necessary, wash
proteins M NaOH for a contact time of 1 to 2 with 1.5 column
hours. Use reversed flow, or volumes of water
wash with 2 to 4 column volumes of a followed by 1
mixture of 1 M NaCl/1 M NaOH for a column volume of 2
contact time of 1 to 2hours. Use M NaCl (not
reversed flow. HIC).Regenerate to
start conditions,
see instructions for
respective media.
Regenerate to start
conditions, see
instructions for
respective media.

6 Maintenance
6.7 Cleaning
Media Contaminants Suggested procedure Follow-up

Lipids Wash with 2 to 4 column volumes of Regenerate to start
0.5 to 2 M NaOH for a contact time of conditions. See
1 to 2 hours. Use reversed flow, or instructions for
wash with 2 to 4 column volumes of respective media.
0.1% to 0.5% non-ionic detergent for a
contact time of 1 to 2 hours. Use
reversed flow, or
wash with 2 to 4 column volumes of
30% isopropanol or 70% ethanol for a
reversed flow.
Sephacryl High Precipitated Wash with 0.5 to 1 column volumes of Wash immediately
Resolution, proteins/lipids. 0.5 M NaOH for a contact time of with at least 2
Sephadex 0.5 to 1 h. Use reversed flow, or column volumes of
G-25 wash with 2 to 4 column volumes of working buffer.
0.5% non-ionic detergent for a contact
time of 1 to 2 hours. Use reversed flow.
Protein A Precipitated or Wash with 1 to 2 column volumes of 6 Wash immediately
Sepharose 4 hydrophobically M Guanidine-HCl for a contact time of with at least 5
Fast Flow, bound 1 to 2 hours. Use reversed flow, or column volumes of
Protein G substances wash with 2 to 4 column volumes of working buffer.
Sepharose 4 0.1% to 0.5% non-ionic detergent for a
Fast Flow contact time of 1 to 2 hours. Use
reversed flow.
Superdex 75 Precipitated Wash with 0.5 to 1 column volume of Wash immediately
and 200 prep proteins/lipids. 0.5 M NaOH for a contact time of with at least 2
grade 0.5 to 1 hour. Use reversed flow, or column volumes of
wash with 2 to 4 column volumes of working buffer.
0.1% to 0.5%non-ionic detergent for a
reversed flow.
After cleaning
After cleaning, restore the column to working conditions using the follow-up proce-
dure. All conditions refer to room temperature unless otherwise stated. If special main-
tenance routines are required, contact your local Cytiva representative for expert
advice. For affinity media, please contact your local Cytiva representative.

6 Maintenance
6.7 Cleaning
Autoclaving
To ensure effective autoclaving it is recommended that the column is completely
disassembled. All plastic components, support net, net, snap plug (BPG 450) and distri-
bution plate (BPG 300 and 450) as well as the wheels and plastic handle caps of the
adjuster nut must be disassembled from the column.
After autoclaving it is recommended that all process contact parts that have not been
autoclaved are replaced with new parts. Essentially seals and nets. Any plastic process
contact parts that are to be reused must be appropriately cleaned prior to reassembly
of the column.
NOTICE
The accessory valves have bodies in PTFE, which can shrink, and
thereby cause leakage after repeated autoclaving. Therefore,
exclude the valves if autoclaving the column.

6 Maintenance
6.8 Storage
6.8 Storage
Bacteriostatic solution
BPG columns packed with media should be stored in the bacteriostatic solution
recommended for the media. Refer to your media instructions for media-specific
recommendations. Refer also to Section 8.2 Chemical resistance, on page 132. Avoid
any solution that may compromise the quality of the media.
Follow the instructions below to apply bacteriostatic solution:
Step Action
1 Clean the column before applying bacteriostatic solution, for example with
0.5M NaOH, or other CIP solution recommended for the media.
2 Rinse the column from all chloride ions.
3 Rinse with purified water after the CIP for at least five column volumes (CV).
4 Apply the bacteriostatic solution and the column is ready to be stored. See
the instructions below for storage of the BPG column.
Store the BPG column

Avoid storage conditions that could expose the column to extreme changes in temper-
ature.
If the column is intended to be stored for longer periods of time, change the storage
solution at regular intervals, for example every 3 months.
Follow the instructions below to store the BPG column:
Step Action
1 Disassemble the column.
2 Perform routine maintenance, i.e., replace O-rings etc.
3 Thoroughly dry all column components.
4 Assemble the column.
5 Seal all inlets and outlets as indicated in wet storage protocol above.
6 Wrap column in plastic to prevent exposure to dust and other airborne

contaminants.
7 Store column at controlled ambient temperatures in a clean and dry envi-

ronment.

6 Maintenance
6.8 Storage
After storage
When the BPG column is going to be taken into operation after storage, follow the
instructions below:
Step Action
1 Disassemble the column.
2 Perform routine maintenance, i.e., replace O-rings etc.
3 Thoroughly dry all column components.
4 Assemble the column.

7 Troubleshooting
7 Troubleshooting
About this chapter

This chapter provides information to operators and service personnel to identify and
correct problems that can occur when operating BPG columns.
If the suggested actions in this guide do not solve the problems, or if the problem is not
covered by the this guide, contact your Cytiva representative for advice.
Air in the column

If air has entered the column, the column can be run at a high velocity in the opposite
direction until de-aired. Place a flow restrictor on the outlet to generate a higher back-
pressure over the bed. However, if only a small amount of air has been trapped between
the column net and the adapter head, it is possible to remove the air by pumping a
solution, with a temperature a few degrees higher than the media, through the column,
in the opposite direction. After reverse pumping, test the column again prior to use,
refer to Section 5.3 Performance evaluation of the column, on page 72.
Troubleshooting guide
Problem Cause Action

High-back pressure The valves between the pump and the Open the valves.
collection vessel are closed.
The valves are blocked. Check that all valves are clean
and free from internal blockage.
The medium is packed too hard. Repack the medium.
The column nets are blocked and there Change nets.
can be air trapped in the net.
The support net is blocked. Clean or change the support
net.
Flow valve selected is not correct. Change flow valve.
Flow cell used not the correct selected. • Select adequate size of flow
cell.
• Check to see if the sample
and collection vessels are at
approximately the same
level as, or above, the pump.

7 Troubleshooting

There are differences in internal diameter Change tubing to the same
between the tubing on the column, the diameter.
tubing from the pump, or anywhere else in
the system.
The pressure gauge is showing faulty Change or calibrate the pres-
values. sure gauge.
Column leakage The end-piece and/or flange is not prop- Disassemble and re-assemble
erly positioned with respect to the tube. the column.
The solvents used are not compatible with Check solvent, see Section 8.2
the O-rings and gaskets. Chemical resistance, on page
132.
The tubing is not compatible with all the Change solvent.
solvents.
The connectors have not the same dimen- Change connectors.
sions.
Air leaks into the column from the suction Check that the connections to
side of the pump. and from the pump are correct
and tightened.
The torque is not correcton the rods or the Check that the correct torque
flange. specification has been used
during column assembly.
Chromatogram not The linear velocity is not as expected. Check sample.
as expected
Back-mixing has occurred anywhere in the Check connections.
column set-up.
The sample could have passed through an Check connections or test
air trap. method.
The adapter is not correctly positioned in • Move the adaptor down 5
close contact with the bed. mm into the bed.
• Disassemble and assemble
the column.
The column packing has not been Check the column packing.
checked.
The column has not been correctly equili- Equilibrate the column.
brated.
Old sample mixed with new. Check sample and sample
handling.

7 Troubleshooting

There have been any changes to the • Check the dilution, filtration,
following: temperature and prepara-
• Preparation of buffers/solutions tion of the sample and
buffers are considered.
• Preparation of system and tubing
• Check all dead volumes were
correctly filled with appro-
priate buffers and sample
before running.
Infected and/or Column bed shows signs of bacterial • CIP.

pyrogen contami- growth.
• Prepare new buffers/sample.
nated column
The starting material is contaminated. Use clean material.
The in going components (such as water, Check material.
serum components in growth media, etc.)
are not ok.
The column is not cleaned. Clean the column, refer to
Recommended CIP procedure,
on page 116.
The NaOH has been removed, in case that Repeat cleaning.
positive LAL tests were seen after a CIP
cycle with NaOH.
Air trapped in the The buffers have not the same tempera- Check the buffers are at the
column ture as the column. same temperature as the
column.
The connections are not correctly fitted Check the connections.
together.
The valves are not shut tight enough. Check the valves and close
them.
The volume of the trap is too small or large. • Fill air trap with buffer.
• Change air trap.

8 Reference information
About this chapter

This chapter contains specifications of BPG. It also contains a list that specifies the
chemical resistance of BPG to different chemicals.
In this chapter
Section See page
8.1 Specifications 125
8.2 Chemical resistance 132
8.3 Abbreviations 135
8.4 Recycling information 136
8.5 Regulatory information 137
8.6 Health and Safety Declaration Form 140

8.1 Specifications
8.1 Specifications
In this section
Section See page
8.1.1 BPG 100 127
8.1.2 BPG 140 128
8.1.3 BPG 200 129
8.1.4 BPG 300 130
8.1.5 BPG 450 131
Column specifications
Bed volumes and bed heights are based on a slurry concentration of 75% and a packing
compression of 15%. The compression is the difference in volume between a sedi-
mented bed and a pressurized bed.
Note: The optimal slurry concentration is close to 50%.

8.1 Specifications
Height
Height (H) used in this section refers to a BPG column with the adapter in its most
upper position.

8.1 Specifications
8.1.1 BPG 100
8.1.1 BPG 100

Parameter Unit BPG 100
Tube height mm 500 750 950
Column inner diameter mm 100
Cross sectional area cm2 78.5
Max column volume l 3.2 5.2 6.2
Max column volume with extension l 6.2 8.2 9.7
Min bed height cm 1 26 46
Bed height1 cm 26 42 55
Bed height with extension2 cm 35 56 74
Bed height with extension3 cm 45 67 N/A
Min bed volume l 0.3 2.0 3.6
Bed volume1 l 2.0 3.2 4.3
Bed volume with extension2 l 2.7 4.3 5.8
Bed volume with extension3 l 3.5 5.2 N/A
Max pressure4 bar(g) 8
Total weight (tare mass) kg 15 16 17
Total weight (tare mass) with extension kg 21 22 23
Adapter weight kg 7
Overall dimension D×W×H cm 48×48×127 48×48×152 48×48×172
Height with extension tube cm 165 190 210
Operating temperature °C 4 to 40
1 Without an extension tube.
2 These bed heights and volumes are achievable using a packing extension during consolidation of the bed (sedimen-
tation). Total amount of 75% slurry must fit into column + extension with adapter.
3 The bed height and volume are achievable when retaining the extension tube during the final packing step. Adapter
must seal at least 5 cm into the column tube to avoid high tensions on glass tube.
4 Use a pressure gauge to monitor the pressure.

8.1 Specifications
8.1.2 BPG 140
8.1.2 BPG 140

Cross sectional area cm2 154
Max column volume l 6.3 10 13
Max column volume with extension l 12 16 19
Bed volume1 l 4.0 6.4 8.4
Adapter weight kg 11

8.1 Specifications
8.1.3 BPG 200
8.1.3 BPG 200

Max column volume l 13 21 27
Bed volume1 l 8.1 13.1 17.2

8.1 Specifications
8.1.4 BPG 300
8.1.4 BPG 300

Bed volume1 l 17.2 28.2 37.1

8.1 Specifications
8.1.5 BPG 450
8.1.5 BPG 450

Bed volume1 l 35.9 60.9 85.9
Max pressure4 bar(g) 2.5

8.2 Chemical resistance

Introduction
This section provides general chemical resistance guidelines for various chemicals
that may be used with BPG columns.
Chemicals to be avoided
In general, the use of the following chemicals in BPG columns should be avoided:
• Extreme oxidizers (such as peroxides)
• Halogenated compounds
• Chlorinated solvents (such as methylene chloride)
• Esters (such as acetates)
• Aromatic hydrocarbons (such as toluene)
• High concentration of strong acids
• Chloroform
• THF
• Toluene
• Methylenechloride
Chemical resistance guide
NOTICE
Only use chemicals listed in the Chemical Resistance information.
The wetted parts of the product may be damaged by chemicals not
listed in the Chemical Resistance information. Contact your Cytiva
representative before using chemicals that are not listed.
Note: Avoid salt in combination with pH below 4.5. Always wash the column with at
least two column volumes of pH-neutral solution between the use of salt
and low pH buffers.
The table below is intended as a guide to the solvent resistance of the materials used in
BPG columns. The information has been compiled from published material from
several sources, not from individual tests on the components of the column. Conse-
quently, the user should use this list only as a guide to the level of chemical resistance
exhibited by each material. It should be noted that the effects of a chemical will be
more severe at higher temperatures and pressures, and that combined effects have
not been taken into consideration.

Substance Concentra- 60–90 days1

tion by
volume
Acetic acid 10% Not recommended to be used with PA nets.
Acetic acid 25% Not recommended to be used with PA nets.
Acetonitrile 5% EPDM rubber changes characteristics. For
repetitive and long term use, use FEP/PFR/
PTFE O-rings and gaskets.
Acetonitrile 50% Change to FEP/PFR/PTFE O-rings and
gaskets.
Polypropylene plastic resistance is fair.
Acetone 50% OK
Cyclohexane 100% Change to FEP/PFR/PTFE O-rings and
gaskets.
Polypropylene plastic resistance is fair.
Ethanol 100% EPDM rubber changes characteristics. For
Ethyl acetate 100% Polypropylene plastic resistance is fair.
Ethylene glycol 50% OK
Glycerol 100% OK
Hexane 100% EPDM rubber changes characteristics. For
Not recommended to be used with PA nets.
Hydrochloric acid 0.1 M Not longer than 4 hours.
Not recommended to be used with PA nets.
Isopropyl alcohol 100% EPDM rubber changes characteristics. For
Methanol 100% OK
Nitric acid 0.1 M Not recommended to be used with PA nets.
n-Propanol 100% OK

Substance Concentra- 60–90 days1

tion by
volume
Sodium chloride 2M Can be used under normal running condi-
tions. Do not use NaCl in storage solutions.
Please note that NaCl can cause corrosion
on stainless steel in acid solutions (pH below
4,5).
Sodium hydroxide 2M OK
Trifluoroacetic 0.1% Not recommended to be used with PA nets.
acid FEP changes characteristics. Thus, use
EPDM rubber.
Detergent 100% OK
Tween™/Tri-n- 1% / 0.3% OK
butyl phosphate
UREA 8M OK
1 The test does not include PVC tubing.
Chemical resistance to sodium

chloride
BPG columns are equipped with wetted components of stainless steel and must there-
fore be appropriately maintained when exposed to NaCl during chromatographic
processes. If a process incorporates the use of NaCl solutions it is essential that the
column be thoroughly rinsed to remove any residual NaCl. It is recommended that a
water rinse of at least five column volumes be used. Water is the preferred rinse solu-
tion due to the solubility properties of NaCl in water verses other possible rinsing solu-
tions like ethanol solution.
The rinsing protocol should use at least five column volumes of purified water to assure
optimal removal of residual chloride ions, which can be corrosive to stainless steel over
time.

8.3 Abbreviations
8.3 Abbreviations
The abbreviations used in this manual are defined as follows:
Abbreviation Description Translation
As Peak asymmetry factor Peak asymmetry factor

CIP Cleaning-in-place Cleaning-in-place
CV Column volume Column volume
EPDM Ethylenepropylenedienemo- Ethylenepropylenedienemo-
nomer rubber nomer rubber
FEP Fluoroethenepropene Fluoroethenepropene
FF Fast Flow Fast Flow
HETP Height equivalent to a theoret- Height equivalent to a theoret-
ical plate ical plate
L Bed height Bed height
N Number of theoretical plates Number of theoretical plates
PA Polyamide Polyamide
PF Pack factor Pack factor
PEEK Polyetheretherketone Polyetheretherketone
PFR Perfluor rubber Perfluor rubber
PTFE Polytetrafluoroethane Polytetrafluoroethane
THF Tetrahydrofurane Tetrahydrofurane
UV Ultra violet Ultra violet
Ve Elution volume Elution volume
WFI Water for injection Water for injection
Wh Peak width at half peak height Peak width at half peak height

8.4 Recycling information
8.4 Recycling information

Decontamination
BPG columns shall be decontaminated before decommissioning and all local regula-
tions shall be followed with regard to scrapping of the equipment.
WARNING
Before decommissioning work is performed on the column, make
sure that:
• the column is empty and depressurized.
• relevant energy sources and process feed are disconnected.
• all process wetted areas are clean and decontaminated.
• the column cannot be accidentally re-energized, refilled or re-
contaminated during maintenance.
WARNING
Decommissioning. Decontaminate the equipment before
decommissioning to ensure that hazardous residues are removed.
Disposal
When taking BPG columns out of service, the different materials must be separated
and recycled according to national and local environmental regulations.
Recycling of hazardous substances

BPG columns may contain hazardous substances. Make sure that there are no
hazardous substances left in the column prior to recycling.

8.5 Regulatory information

Introduction
This section lists the regulations and standards that apply to the product.
In this section
Section See page
8.5.1 Contact information 138
8.5.2 European Union and European Economic Area 139

8.5.1 Contact information
8.5.1 Contact information
Contact information for support

To find local contact information for support and sending troubleshooting reports, visit
cytiva.com/contact.
Manufacturing information
The table below summarizes the required manufacturing information.
Requirement Information
Name and address of manufacturer Cytiva Sweden AB
Björkgatan 30
SE 751 84 Uppsala
Sweden
Telephone number of manufacturer + 46 771 400 600

8.5.2 European Union and European Economic Area
8.5.2 European Union and European Economic Area
Introduction
This section describes regulatory information for the European Union and European
Economic Area that applies to the equipment.
Conformity with EU Directives

See the EU Declaration of Conformity for the directives and regulations that apply for
the CE marking.
If not included with the product, a copy of the EU Declaration of Conformity is available
on request.
CE marking
The CE marking and the corresponding EU Declaration of Conformity is valid for the
instrument when it is:
• used according to the Operating Instructions or user manuals, and
• used in the same state as it was delivered, except for alterations described in the
Operating Instructions or user manuals.

8.6 Health and Safety Declaration Form

On site service
On Site Service Health &

Safety Declaration Form
Service Ticket #:
To make the mutual protection and safety of Cytiva service personnel and our customers, all equipment and work areas must be
clean and free of any hazardous contaminants before a Service Engineer starts a repair. To avoid delays in the servicing of your
equipment, complete this checklist and present it to the Service Engineer upon arrival. Equipment and/or work areas
not sufficiently cleaned, accessible and safe for an engineer may lead to delays in servicing the equipment and could be subject to
additional charges.
Review the actions below and answer “Yes” or “No”.

Yes No
Provide explanation for any “No” answers in box below.
Instrument has been cleaned of hazardous substances.

Rinse tubing or piping, wipe down scanner surfaces, or otherwise make sure removal of any dangerous residue.
Make sure the area around the instrument is clean. If radioactivity has been used, perform a wipe test or other
suitable survey.
Adequate space and clearance is provided to allow safe access for instrument service, repair or
installation. In some cases this may require customer to move equipment from normal operating location
prior to Cytiva arrival.
Consumables, such as columns or gels, have been removed or isolated from the instrument and from
any area that may impede access to the instrument .
All buffer / waste vessels are labeled.

Excess containers have been removed from the area to provide access.
Provide
explanation
for any “No”
answers here:
Equipment type / Product No: Serial No:
I hereby confirm that the equipment specified above has been cleaned to remove any hazardous substances and that the area
has been made safe and accessible.
Name: Company or institution:
Position or
Date (YYYY/MM/DD):
job title:
Signed:
Cytiva and the Drop logo are trademarks of Global Life Sciences IP Holdco LLC or an affiliate.
© 2020 Cytiva.
All goods and services are sold subject to the terms and conditions of sale of the supplying company operating
within the Cytiva business. A copy of those terms and conditions is available on request. Contact your local Cytiva
representative for the most current information.
For local office contact information, visit cytiva.com/contact.

28980026 AD 04/2020

Product return or servicing
Health & Safety Declaration Form

for Product Return or Servicing
Return authorization and/or
number: Service Ticket/Request:
To make sure the mutual protection and safety of Cytiva personnel, our customers, transportation personnel and our environment,
all equipment must be clean and free of any hazardous contaminants before shipping to Cytiva. To avoid delays in the processing of
your equipment, complete this checklist and include it with your return.
1. Note that items will NOT be accepted for servicing or return without this form
2. Equipment which is not sufficiently cleaned prior to return to Cytiva may lead to delays in servicing the equipment and
could be subject to additional charges
3. Visible contamination will be assumed hazardous and additional cleaning and decontamination charges will be applied
Yes No Specify if the equipment has been in contact with any of the following:
Radioactivity (specify)
Infectious or hazardous biological substances (specify)
Other Hazardous Chemicals (specify)

Equipment must be decontaminated prior to service / return. Provide a telephone number where Cytiva can contact
you for additional information concerning the system / equipment.
Telephone No:
Liquid and/or gas in equipment is: Water
Ethanol
None, empty
Argon, Helium, Nitrogen
Liquid Nitrogen
Other, specify
Equipment type / Product No: Serial No:
I hereby confirm that the equipment specified above has been cleaned to remove any hazardous substances and that
the area has been made safe and accessible.
Company or
Name:
institution:
Position or job title: Date (YYYY/MM/DD)
Signed:
Cytiva and the Drop logo are trademarks of Global Life Sciences IP Holdco LLC or an affiliate. To receive a return authorization number
© 2020 Cytiva. or service number, call local
All goods and services are sold subject to the terms and conditions of sale of the supplying company operating technical support or customer service.
within the Cytiva business. A copy of those terms and conditions is available on request. Contact your local Cytiva
representative for the most current information.
For local office contact information, visit cytiva.com/contact.

28980027 AD 04/2020

Index
Index
A Cleaning, 37, 115
agents, 115
Adapter, 76, 81, 82, 101, 106 Cleaning-in-place, 116
assembly, 106 Column, 20, 37, 39, 59, 72, 79, 87,
disassembly, 101 90, 94, 119, 123
moving, 76 assembly, 59
net, 81 blockage, 79
remove, 82 change O-ring, 94
Air, 54, 121 contaminated, 123
remove from column, 121 disassembly, 90
remove from top of adapter, leveling, 39
54 material, 20
Air trapped, 123 moving, 37
As, 72 nets, 87
ATEX directive, 21 performance evaluation, 72
Autoclaving, 118 storage, 119
Column leakage, 122
B Columns, 20, 34
floor load, 34
Bacteriostatic solution, 119 net, 20
Bed height, 50 space requirements, 34
BPG 100-140-200 columns illus- Compression factor, 50
trations, 22 Connections, 47
BPG 300 columns illustrations, 25 arrange, 47
BPG 450 columns illustrations, 28
BPG columns, 6, 19, 20, 31, 125 D
accessories, 31
design pressure, 19 Decontamination, 136
intended use, 6 Disposal, 136
material, 20 general instructions, 136
specifications, 125 Drain, 43
tubing, 20 symbol, 43
volumes, 19
E
C
Emergency, 17
Calculation, 73, 74 restart, 17
assymetry, 74 shutdown, 17
HETP, 73 Emergency stop, 17
CE, 139 End-piece items, 87
conformity, 139 Environment, 34
marking, 139 Evaluation, 72
Chemical resistance, 132 column performance, 72
Chromatogram, 73, 122 Explosive atmosphere, 21
CIP, 116 Explosive environment, 10
protocol, 116 precautions, 10

Index
Extension tube, 31, 99 O

fitting, 99
O-rings, 85, 92, 94
F replacement, 92
replacement adapter, 85
Flammable liquids, 10 replacement column, 94
precautions, 10 On site service, 140
Flow rate, 71 Operation, 12
Four-port valve, 44, 98 precautions, 12
positions, 44
replace sealing washer, 98 P
G Packing column, 62, 67, 69
first step, 62
Grounding, 40 remove buffer from adapter,
Grounding kit, 41 69
second step, 67
H Packing extension tube, 31, 48
Packing flow rate, 71
HETP, 72, 73 plot the result, 71
High-back pressure, 121 Passivation, 78
Personal protection, 11
I Pressure flow curve, 71
Pressure gauge, 43
Important user information, 6 Product return or servicing, 141
Installing, 11 Purpose of this manual, 5
precautions, 11
R
L
Recycling, 136
Leakage test, 113 hazardous substance, 136
Location, 47 Regulatory information, 137
Remove buffer, 69
M Replacement, 80
tools, 80
Maintenance, 13, 77 Restart, 17
precautions, 13
Manufacturing information, 138 S
Moving, 11
precautions, 11 Safety notices, 6
Safety precautions, 9
N Service, 77
frequency, 77
Net, 48, 85 Service agreements, 77
adapter, 85 Setup, 43
end-piece, 48 Slurry, 50, 52
replacement, 85 concentration, 50
Nets, 20, 93 preparation, 52
column, 93 Slurry volume, 50
Notes and tips, 7 Storage, 119
System operation, 12
precautions, 12

Index
T
Tools, 36, 80
replacement, 80
unpacking, 36
Torque wrench, 88
Transport, 35
crate, 35
Troubleshooting, 121
U
Unpacking, 36
instructions, 36
W
Wheel, 38
lock, 38

Page intentionally left blank
cytiva.com
Cytiva and the Drop logo are trademarks of Global Life Sciences IP Holdco LLC or an affiliate.
BioProcess, Sephacryl, Sephadex, Sepharose, and Superdex are trademarks of Global Life Sciences Solutions USA LLC or an affiliate
doing business as Cytiva.
Tween is a registered trademark of Croda Group of Companies
All other third-party trademarks are the property of their respective owners.
© 2020 Cytiva
All goods and services are sold subject to the terms and conditions of sale of the supplying company operating within the Cytiva
business. A copy of those terms and conditions is available on request. Contact your local Cytiva representative for the most current
information.
For local office contact information, visit cytiva.com/contact
29193223 AB V:5 12/2020

29193223

Uploaded by

Copyright:

Available Formats

29193223

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

29193223

Uploaded by

Copyright:

Available Formats

BPG Columns

2 Safety instructions ............................................................................................. 8

3 Column descriptions .......................................................................................... 18

2 BPG Columns Operating Instructions 29193223 AB

6.8 Storage .............................................................................................................................................................. 119

7 Troubleshooting ................................................................................................ 121

8 Reference information ....................................................................................... 124

Index ........................................................................................................................... 142

BPG Columns Operating Instructions 29193223 AB 3

About this chapter

Section See page

1.1 About this manual 5

1.2 Important user information 6

4 BPG Columns Operating Instructions 29193223 AB

1.1 About this manual

Scope of this document

BPG Columns Operating Instructions 29193223 AB 5

1.2 Important user information

Intended use of the product

6 BPG Columns Operating Instructions 29193223 AB

Notes and tips

BPG Columns Operating Instructions 29193223 AB 7

About this chapter

Section See page

2.1 Safety precautions 9

2.2 Symbols and abbreviations 15

2.3 Emergency procedures 17

8 BPG Columns Operating Instructions 29193223 AB

2.1 Safety precautions

BPG Columns Operating Instructions 29193223 AB 9

Flammable liquids and explosive

10 BPG Columns Operating Instructions 29193223 AB

Installing and moving

BPG Columns Operating Instructions 29193223 AB 11

12 BPG Columns Operating Instructions 29193223 AB

BPG Columns Operating Instructions 29193223 AB 13

14 BPG Columns Operating Instructions 29193223 AB

2.2 Symbols and abbreviations

Description of symbols on the system

Label text Description

The equipment complies with applicable European direc-

BPG Columns Operating Instructions 29193223 AB 15

Placement of system label

16 BPG Columns Operating Instructions 29193223 AB

2.3 Emergency procedures

2 If an EMERGENCY STOP button was used to shutdown the equipment

3 Reduce pump settings prior to reactivation to avoid immediate high pres-

4 Restart the equipment connected to the column.

BPG Columns Operating Instructions 29193223 AB 17

About this chapter

Section See page

3.2 BPG 100, 140 and 200 columns 22

3.3 BPG 300 column 25

3.4 BPG 450 column 28

18 BPG Columns Operating Instructions 29193223 AB

BPG Columns Operating Instructions 29193223 AB 19

Adapter and rods

20 BPG Columns Operating Instructions 29193223 AB

Definition of the zone A place in which an explosive atmosphere consisting

BPG Columns Operating Instructions 29193223 AB 21