PRAXSA

3M Solutions for Biopharmaceutical Process Development,

Manufacturing and Process Monitoring

www.praxsasrl.com info@praxsasrl.com
 3M Is The Innovation Company That Makes Progress Possible

 We create transformational products and solutions that enable customer success and improve
people's lives around the world.

 We utilize a collaborative, high-energy approach to solve the toughest problems across

industries and markets by:
 Constantly exchanging and building on each other's ideas
 Uncovering new connections between seemingly unrelated markets and more than 50
diverse technology platforms
 Fostering a culture of intellectual curiosity and creativity that pushes boundaries

At 3M, we are advancing the global biopharmaceutical industry by helping to build better
and more efficient manufacturing processes, improving product safety by providing tools for
monitoring and tracking, and reducing energy usage with our technologies. The Life Sciences
Process Technologies business unit of 3M Purification Inc. provides cutting edge technologies to
address clarification, filtration and purification needs of the global biopharmaceutical industry.

Figure 1: The Biopharmaceutical Production and Downstream Purification Train

Media
Buffer
Bioreactor Cell Culture Clarification Pre-Purification Protein-A Capture Low pH Hold Column Protection HIC Column AEX Flow-Through Virus Filter UF/DF Final Filtration

Air Filter Air Filter Buffer Filtration / In-process Filtration Buffer Filtration / In-process Filtration
In Out

2 3
PRAXSA

3M Technologies
At 3M, our technology, products and innovation reflect what we do for our customers every day:
advance, enhance and improve their products and processes to enable their success.
3M Purification's dedicated technical services and laboratory personnel help solve customer’s most
arduous separations problems. Our engineers work to provide solutions that reduce the overall cost
Viral Based
of ownership. Our researchers are constantly working on breakthroughs that make new separations
Therapeutics
platforms possible.
Every day our products are used by researchers, process developers and manufacturing personnel for
critical filtration, separation and process monitoring steps in the biopharmaceutical industry.

Diagnostics Reagents &

S
Serum Filtration

Bacterial and Yeast

Based Cell Culture,
e.g. Insulin

Biopharmaceutical Separations Applications

Biopharmaceutical refers to biologically active therapeutic
and diagnostic proteins that are expressed by mammalian,
insect, yeast or bacterial cells. Such drugs can be classified
into: monoclonal antibodies, growth factors, hormones,
cytokines, fusion proteins, and therapeutic enzymes. Viral
based therapeutics are poised to grow rapidly and hold great
promises for disease treatment in the future. The manufacturing
Follow-on Biologics —
Biosimilars & Bio-betters process of diagnostic reagents and sera contains many
biopharmaceutical separations applications.
Filtration and purification plays an essential role in
manufacturing of biopharmaceutical drugs. 3M offers a range
of filtration, purification and process monitoring technologies
Mammalian Cell Culture — that can be used in both upstream and downstream steps in
e.g. Monoclonal Antibodies, Fusion Proteins every scale of biopharmaceutical manufacturing.

4 5
www.praxsasrl.com info@praxsasrl.com
The Bioreactor is at the heart of a biopharmaceutical manufacturing      Automated
Integrity Tester
Surface
Cleanliness
Air Filtration Vent Air Filtration In Process Filtration Of Filtration Of
process. For the bioreactor to work at maximal efficiency, filters Storage Media Nutrients/Buffers For Housings And Monitoring
Components Connections
used for air and media filtration need to completely remove foreign
High LRV removal of Reliable removal of Portable handheld Real-time monitoring
microorganisms. Scope of
application†
Retention of bacteria and aerosolized
bacteriophage
Short term storage of
media, buffers etc.
mycoplasma for media bacteria from nutrients integrity tester to mea- of microbial / protein
components / buffers sure pressure decay. residues from surfaces
Filter Pore
0.2 µm Not Applicable 0.1 µm 0.2 µm Not Applicable Not Applicable
Size

LifeASSURE™ PFS LifeASSURE™ PSA MiniCheck™

3M™ Single-Use 3M CleanTrace™
LifeASSURE™ PDA
Biocontainer

3M
Products

Proprietary Multi-layer PES membrane Filter

Electronic Instrument
Materials PTFE membrane Filter Film Nylon membrane Filter (Sterile and Gamma Electronic Instrument
with consumables
(LDPE contact layer) Compatible)

Validated for sterilizing performance using a liquid

bacteria B. diminuta (ATCC 19146) at challenge Microprocessor driven, 3M Clean-Trace™
Detailed validation Absolute retention
levels of a minimum of 107 CFU/cm² for reliable Absolute retention of programmable system Surface and Water ATP
package outlining of Acholeplasma
Validation sterilizing performance in wet or dry conditions B. diminuta (ATCC (with available printer) tests help assess stan-
extractables including laidlawii, ATCC 23206
of operating 19146) at challenge for determining leak dards of hygiene and
LC/GC-MS data with at challenge levels of
properties levels of > a minimum integrity of closed pres- cleaning procedures by
various buffers and > a minimum of 107
Complete bacteriophage retention using aerosol of 107 CFU/cm² surized systems (e.g. measuring the amount
chemicals CFU/cm²
challenge test methods Filter and housings) ATP or proteins

Bacterial Fermentation e.g. Escherichia coli †

Developed in the late 1970s, the bacterial expression system, such as the E. coli system, is used to produce many important therapeutic
recombinant proteins. The popularity and dominance of the bacterial system has continued until today because of its advantage in speed,
simplicity and cost. The challenge of using this system is in downstream purification. A homogenization step after harvest is often needed
because most of the expressed proteins are located in inclusion bodies rather than secreted outside the cells. This step increases the level
of lipids, host cell proteins, DNA and endotoxins along with the target protein; hence resulting solutions from bacterial harvest are difficult to
purify.

Mammalian Fermentation e.g. CHO Cell†

Mammalian cells (e.g., CHO [Chinese Hamster Ovary] Cells) are typically used to produce monoclonal antibodies. Monoclonal antibodies are
derived from one single clone and thus are identical in structure. Today over 30 monoclonal antibodies have been approved in the U.S. with
indications for a variety of difficult to treat diseases such as autoimmune diseases, cancers, infectious diseases, etc. Clarification involves
separation of the cells from bulk media. Cell density (> typical 5 x 106 cells/mL) and cell viability affect the depth filtration step. In general,
Figure 2: 3M Purification Filters and Biocotaniers for use with Bioreactors mammalian cell cultures are easier to filter than bacterial cell culture harvests.

6 7
PRAXSA

Figure 3: Cell Clarification Set-Up† Figure 5: Detailed View of Cell Culture Clarification†

3M offers the most Centrifuge

comprehensive portfolio Method
of depth filters for cell
culture clarification in the
biopharmaceutical industry.
Zeta Plus™ depth filtration Bacterial Cell Culture Zeta Plus 60ZA05A or 90ZA08A Media
technology, in cartridge Mammalian Cell Culture Zeta Plus 60ZA05A Media
systems and sheets, play
an important role in the Depth Filtration
clarification of cell-derived Method
protein therapeutic products
around the world. 3M is
recognized as a market leader
in depth filter technology.

Bacterial Cell Culture Zeta Plus 60SP02A Media

Mammalian Cell Culture Zeta Plus 60SP02A or 90ZA08A Media

Coarse Clarification

10.0
Polishing

Table 1: Depth Filter Media Recommendations†

Contaminant Size (Microns)

Fluid
Feed Composition Recommendation

Sediments
Turbidity Ultrafine
Filtration

Yeast
Whole cells, hard particles • Centrifuge
(density gradient ). > 300 NTU • TFF 1.0

First clarification. • Open pore depth filter (05 or 10 SP) Table 2: Range of Media Adsorption Properties
100-300 Designation Media Surface Characteristic
Colloidal, Cell Debris • Medium Pore Depth filter 30-60 SP
NTU
ZA Strong Anion-Exchange
Colloidal, Small Particulates 20-100 NTU • 60SP or 60ZA or 90 ZA

Bacteria

Hazes
SP Medium Anion-Exchange
Fine particulates, colloidal
< 10 NTU • 90 or 120 ZA HP Weak Anion-Exchange High Wet Strength 0.1
0.1
(Final Polishing) 120 90 70 60 50 30 10 05
Intra-cellular, requires cell ZC Activated Carbon for color / organic adsorption Zeta Plus™ Grades
> 300 NTU • Centrifuge
breakage – First Clarification Step Figure 4: Zeta Plus™ Depth Filter Family DELI Activated Silica for adsorption of hydrophobic moieties Figure 6: Wide Choice of Media Grades
8 9
www.praxsasrl.com info@praxsasrl.com
Traditional Solutions for Cell Culture Clarification Single-Use Solutions for Cell Culture Clarification
Based on our more than 30 years of technical expertise in filtration and separation products for the 3M offers a complete package of single-use systems for cell culture clarification applications for
biopharmaceutical industry, we offer: biopharmaceutical customers. Single-use Zeta Plus™ Encapsulated depth filter clarification solutions are
The broadest portfolio of lenticular depth filter media in the industry available in scaleable capsule formats from R&D to process development to pilot / clinical production to
 Complex custom engineered systems commercial and large scale production. In addition, we also offer customized solutions and accessories, such
Single-use storage solutions and customized connector sets as tubing connectors and staging carts to round out a complete package.

 Tools for integrity testing of filter systems, handheld pressure decay measurement systems
16EZC
DL — 56 m²
SL — 87.5 m²
 Range of sterile and gamma compatible 0.2 μm PES membrane capsules

Capacity
16EZB
DL — 11.2 m²
SL — 17.5 m²

16EZA
DL — 1.6 m²
DL = Double Layer SL — 2.5 m²
SL = Single Layer

Zeta Plus™  Complex Custom  E1020

1020 cm² 2,000 - 25, 000 L

Lenticular Cartridges Engineered Solutions 200 - 5,000 L

E0170 170 cm²
25 - 200 L
E0340 340 cm²
5 -25 L
BC25
25 cm²
1-5L

0.5 - 1 L Process Volume

Figure 7: Scalable Single-Use Solutions from 0.5 to 25,000 liters

Accessories
Custom Single-use Tubing Connectors for Zeta Plus™ Encapsulated Systems Staging Carts – For Loading Capsules

 
INLET &/OR OUTLET 314
1"/ 1 1/2" CONN.
(TYP.)

 Tools for 72° (TYP.)

18°

312
54°
1
374
54°
98°
48°

1
374

3M™ Single-Use Integrity Testing of LifeASSURE™ PDA 111

16
72°
18°

(TYP.)

Biocontainers Filtering System 0.2 µm Sterile Capsules 67

8
(TYP.)
R8
297
8
R8

291

30
2

11
27
16
(TYP.) 30 9
16
(TYP.)

FRONT VIEW FRONT VIEW

10 11
PRAXSA

Zeta Plus™ Encapsulated Systems are designed to make cell Advantages of The Zeta Plus™ Encapsulated system
clarification by depth filtration fast, easy and clean. 3M offers three
 Simplifies the operation of depth filtration step
models of Zeta Plus Encapsulated Holders — 16EZA, 16EZB and
 Full utilization of the filter media, small footprint Zeta Plus™ Encapsulated System (Model 16EZB) Zeta Plus™ Encapsulated Multi-Round System (Model # 16EZC)
16EZC — as a convenient single-use depth filter system for cell
culture clarification. Both the Single Round (Model #16EZB) and during filtration For up to 5,000 Liters For up to 25,000 Liters
Multi-Round (Model #16EZC) can be pivoted between horizontal  Avoids spills on the manufacturing floor
and vertical positions, allowing for convenient loading and  Ergonomic and saves labor
unloading, minimal footprint during filtration, minimal fluid spills
during unloading, and full utilization of the filter media. The pilot
scale system, 16EZA, uses up to 3.2 m² of depth filter media and is
not designed to pivot.
Model 16EZB
Model 16EZC
At the heart of the Encapsulated Zeta Plus system is the uniquely designed Depth Filter Capsule.
Translucent plastic shell allows for
A cam structure allows reliable
easy liquid level detection
capsule-to-capsule connection

Upstream hold-up volume is

reduced to minimum

Handles designed for easy

loading and unloading Solid core design eliminates the need for Replacing a centrifuge in CHO cell harvest process†
central post and stainless steel band
The Zeta Plus™ Encapsulated single-use system has been used to replace a centrifuge and a conventional single layer depth filter in an existing pro-
cess for CHO cell culture clarification at a major biopharmaceutical manufacturer.
Justification: Operator Feedback:
Table 3: Zeta Plus™ Encapsulated System Specifications
During harvest runs, the continuously stacked • Minimal residual liquid was observed during system break-down.
16EZB 16EZC disc centrifuge broke down periodically, caus- • Zeta Plus Encapsulated system was easy and convenient to set up.
Dimensions (nominal)
1.0 m x 0.5 m x 2.2 m 2.9 m x 2.5 m x 2.2 m ing maintenance and operational bottle necks. • Single-use depth filtration significantly reduced cleaning time for equipment and the process suite.
(39.4" x 19.7" x 86.6") (114.2" x 98.4" x 86.6") Going from two-unit operations to one and • No CIP or cleaning validation studies were required.
Carriage Array Options 1 rack of capsules 3 or 5 racks of capsules converting a hard plumbed system into • Further, the cycle times were shortened thus reducing manufacturing costs.
Loading per Rack 1 - 7 capsules 7 capsules a single-use process results in significant
Can load just one rack and savings.
Can plumb for two stage depth filtra-
leave others empty. Can
Flexibility tion in the same system (maximum Solution:
plumb for two stage depth
number of capsules - 6)
filtration
3M’s Zeta Plus EXT media grade 60SP02A in
Manual (interlocked by PLC to
Torque Limiter Manual an encapsulated format
ensure capsules are secure)
Indexing and Pivoting to Automated with torsion limited Details:
No indexing. Pivot by manual gear.
Vertical Position electric motors.
Zeta Plus™ Encapsulated Capsule Zeta Plus™ Encapsulated Capsule 11 m² of Zeta Plus Encapsulated module
With Standard Polycarbonate Shells With Alkaline Resistant* Control Manual - All mechanical parts PLC controlled
processed 1,000 liter batch and differential
Polyphenylene / Polystyrene Shells Filter Area
Double Layer Media up to 11 m² up to 55 m²
pressure across the downstream 0.2 μm
Single Layer Media up to 17.5 m² up to 87.5 m² membrane was < 2 psid (0.15 bar) throughout
the run
* Based on testing with 1M NaOH and 5% NaClO (Bleach). See Chemical Com-
patibility Guide (70-0202-2023-5/LITPHG03) for more information. 12 13
www.praxsasrl.com info@praxsasrl.com
Exploiting The Charge
Effect Of Depth Filters†
Depth filters are made of cellulose fibers,
a filter aid (e.g. Perlite) and binding resins
that impart a charge to the filtration
matrix. 3M offers a range of depth filter matrices that have strong anion 1.4 Endotoxin Removal†
100
exchange (ZA grade media) to weak anion exchange (SP grade media) 1.2
Endotoxins are cell wall components of gram-negative bacteria consisting of
60ZA - Strong AEX
characteristics. 1
60SP - Weak AEX lipopolysaccharides that can cause a pyrogenic response in a parenteral formulation.

(mg Metanil Yellow

98

Charge Capacity
When a protein therapeutic is made in a bacterial expression systems, such as E. coli,
0.8

Dye/cm^2)
Monoclonal antibodies (mAbs) typically have isoelectric points (IEP or pI)

Endotoxin Removal (%)

0.6
that vary from pH of 4.5 to 8.51. If the pI of the mAb is greater than the downstream processes are required to remove these endotoxins below the threshold level. 96
0.4
pI of the HCP/DNA contaminants, these contaminants can be removed Fortunately, because of its high negative charge, endotoxin can be removed using media
0.2
that have anion-exchange characteristics. Graph 2 shows, endotoxin removal of various
94
Sterile Water FŽƌ/ŶũĞĐƟŽŶ
by exploiting the operating pH and choosing the optimal depth filter resin 0
Phosphate buīer pH 7.0
Phosphate buīer pH 8.5

system. It is important that the feed stream be relatively free of colloidal 3.5 5 6 7 9 Zeta Plus grades filters3. 92
pH
particles as they are also adsorbed by the charged depth filter. Graph 1 Zeta Plus 120ZA10A

Graph 1 above shows charge capacity of strong anion exchange and weak
loading 200 L/m2

25 cm ID Pooled
DNA Removal 90
60ZA05A 90ZA08A 120ZA08A
Perfusion Chromatography 3.68 m2 Depth
Table 4: Percent Capture of DNA by Charged Depth Filter Media Protein A Eluate for Zeta Plus Media Grades
anion exchange resin as a function of pH. The graph shows Zeta Plus ZA
Harvest Skid Filter
Column hc DNA Downstream processes
(strong anion exchange [AEX]) maintains the charge at higher pH, while Zeta Plus™ 90SP Grade Media % Removal of DNA Sample every 30 min are required to reduce Graph 2
Zeta Plus SP (weak AEX) has reduced charge capacity as pH increases. Lot Number pH 7.4 pH 9.0 the residual host cell DNA to levels less than 100 pg / dose of the final
DNA Removal at a pH 7.4 and 9.0 for Zeta Plus SP grade filters is shown 24022 99.9 29
Large Scale hcDNA Clearance in Harvest protein therapeutic formulation per safety requirements of US FDA.
in Table 4. 24107 100.0 45 100,000 Relatively high levels of DNA are seen in perfusion harvest and large
Feed: 55,968 ng/mg
23443 99.8 68 amount of DNA co-elutes with the protein from the Protein-A column.
Zeta Plus SP grade depth filters have lower AEX capacity at pH 9 Conclusions:
10,000 -DNA level reduced by > 300 fold in Įltered harvest The challenge for the downstream purification process is to consistently
compared to pH 7.4. -DNA removal more eīecƟve at large scale
Pooled Average:
reduce DNA levels in the final product. DNA can bind nonspecifically to

ng DNA / mg Protein
Surface Retention
1,000 183 ng/mg
the backbone of Protein A column. Due to its phosphate groups, DNA is
Charge
highly negatively charged at physiological pH and thus is well suited to be
Adsorption

+ = 100
removed by binding to AEX ligands4.
10
Zeta Plus™ depth filters can be used for DNA removal. Graph 3 shows
Depth Filter

Charged depth filter media ...featured in Zeta Plus ... acts like a shallow
Zeta Plus ZA media’s DNA removal properties.
1
such as Zeta Plus ZA or DELI media... low hold-up depth filter formats ... chromatography column

Figure 8: Liters Harvest / mРĞƉƚŚ&ŝůƚĞƌ

Reference: Reduction of Host Cell DNA in Mammalian Cell Culture Using Charged Filters at Protein-A Capture for Monoclonal Antibod
John Wesner Associate Scientist Development Pilot Plant Recovery Operations, Centocor R&D in BPI Korea 2008

Host Cell Protein Removal†

Graph 3
Host Cell Proteins (HCPs) are undesired components in down stream protein purification processes. HCPs are known to foul Protein
A resins, thereby impacting the operation of this important purification step. In addition, the presence of high levels of HCP may lead
to protein precipitation, or aggregation, either before or after the Protein A column. Low pH viral inactivation is particularly prone to Product Recommendations For Pre-Purification Applications†
protein precipitation. Protein aggregates can sporadically plug the sterile filters. Single-Use Format Effective Area
Anion Exchange
Choice of Media BC25 25 cm²
160 Figure 9 shows, the flow Functionality
Scale-Up Capsule 170 cm², 340 cm², 1020 cm²
schematic and levels of
OD 410 nm*mL (Measure of HCP)

140 LA, LP, SA & SP Grades Tertiary Amine

Single Stage Depth Filtration
Two Stage Depth Filtration Small Zeta Plus™ Encapsulated 0.23 m²
120 HCP removal using Zeta ZA Grade Quarternary Amine
Cell Culture ~ 400 L Zeta Plus 90ZA grade media 100 Large Zeta Plus™ Encapsulated 1.6 m²
Plus™ 90ZA depth filter
First Depth Second Depth 80
Filter Pass Filter Pass 60
media2. Application Notes
50 LMH Filtration Grade Pore Rating
Hold 40 • Flow Rate - 50 LMH
Tank 20
30 5.0 - 1.5 µm HCP • Typical Depth Filter Capacity 150-200 L/m²
Frac.1 Frac.1 60 0.8 - 0.2 µm • Two stages work better than single
0
Fract2 Fract2 0 20 40 60 80 100 120 140 160
Volumetric Loading (L/m^2) 90 0.5 - 0.1 µm • Flow rate -- 100 LMH
Frac3 Frac3 DNA Removal
• At Neutral and low pH DNA has a net negative charge
t
Depth
filter
NFEJB
HSBEF
90ZA
used
in
first
and
second
pass 120 0.3 - 0.1 µm
t
Filtrate
fractions
were
processed
with
a
small
scale
Protein
A

• Removal varies as a function of endotoxin challenge levels,
Endotoxin Removal
Figure 9: HCP Removal by Charged Depth Filters column
to
assess
turbidity
and
HCP
elution type of solution (WFI, buffer etc.) and operating pH

14 15
PRAXSA

Particulate Removal prior to Chromatography Columns†

In biopharmaceutical downstream processes, protein refolding is effected by adjusting the pH and salt concentration of Zeta Plus™ Media and Selection Recommendations†
the feed solution. However, an undesirable by-product of this refolding process is the resultant particulate precipitates,
or aggregates. Presence of particulates in chromatography feeds can result in plugging, fouling or loss of performance  Zeta Plus™ depth filters are used to remove particulates prior to chromatography columns
of the column. 3M's depth filter media can be used to remove particulates prior to Cation Exchange (CEX) or Anion-  Zeta Plus DEL series filter media contains activated silica that selectively adsorbs lipids, deter-
Exchange (AEX) column steps. gents, anti-foams that foul chromatography resins ultrafiltration (UF) systems and sterile mem-
brane filters
Tissue Pathway
Inhibitor Factor
(TFPI) expressed
in E.Coli
Zeta Plus Media Choices and Selection Recommendations
Recovery of Two different types of Zeta Plus DEL series filter media are available to suit varying application
Inclusion Body
requirements. The chart below serves as a guideline for selecting the appropriate media type:
Refolding of Pro
tein
(Leads to partic
ulates)

Zeta Plus™ 60
LP
Depth Filter Me
dia
0.2 μm
Membrane
Cation Exchange
Column

Figure 11: Particulate Removal Prior to CEX5

Type Lipid Optimized for Low Aluminum Optimized for

Reduction Capacity Extractable Levels Sensitive LAL* Test Procedures
Improving Hydrophobic Interaction DELI Intermediate No No
Chromatography (HIC) Column Table 5: HIC Performance with and without Zeta Plus DEL Series Media ™ DELP High Yes Yes

Performance† Pre-treatment (Loading volumes were adjusted to contain the same Virus-like Particle (VLP)* per *Limulus Amoebocyte Lysate
cycle and equivalent percentage lipid amounts were calculated)
Zeta Plus™ DEL series filters combine high lipid removal Feed to HIC Column Pretreatment
with cellulose-based depth filtration and can remove
No Filter Zeta Plus™DEL Anti-foam removal from cell-culture solutions†
hydrophobic components and particulates such as lipids, Series Filters
protein aggregates and cell debris in a single step prior
Lipid in the Feed 100% 43% Application Details
to a HIC unit operation6. Table 5 shows the effect of pre-
Loading 33% 80% • Anti-foam agents (e.g. Sigma Aldrich Antifoam A, silicone polymer, Dow ‘Antifoam C’)
treatment with Zeta Plus DEL series filter prior to an HIC
First Cycle Elution 93% 90% • Typical concentrations 1-50 ppm
column and the resultant improvement in performance. To
VLP Amounts Yield 31% 72% • Not cleared in chromatography steps, co-elutes
Further
in HIC Column Loading 62% 87% • Anti-foams foul downstream chromatography columns, UF and sterile membranes
Processing
40th Cycle Elution 29% 43% 3M Solution
Yield 18% 37% • Zeta Plus Series Filter Media: contains hydrophobic/ lipid selective adsorptive media,
cationic resin and cellulose
*Lipid foulant interactions during chromatographic purification of virus like particles from Saccharomyces Cell Culture Zeta Plus™
Cell Culture Zeta Plus™ HIC Column
Harvest Delipid Media
cerevisiae , Ph D. Thesis, Jing Chin, 2010, University College, London Harvest DEL Series Media
Figure 12: Improving HIC Column Performance
16 17
www.praxsasrl.com info@praxsasrl.com
PES — The Preferred Membrane For
Biopharmaceutical Applications Table 7: 3M Membrane Choices For Sterile Filtration Applications†
• Polyethersulfone (PES) membranes offer higher flow rates
• Broad chemical compatibility Membrane 3M Product Pore Size Applications Additional Features
• Wide pH range: Same filter for acid and basic buffers Polyethersulfone LifeASSURE™ 0.6 / 0.2 µm • Final Filtration • Capsules in gamma pre-sterilized
(PES) PDA Series Two Layers • In-process Filtration version available
• Low protein binding and adsorption Filters • Filtration of chromatography pools • Sterilizing Grade Filter
• Post-depth filter sterile filter • Rated to remove 107 B. diminuta
• Buffer filtration per ASTM ASTM F838-05 tests
Sterilizing grade filters are widely used in many downstream processing • 100% integrity tested prior to
steps. In addition to the sterile filtration prior to the final filling step, release
sterilizing filters are used widely in many intermediate steps to reduce Polyethersulfone LifeASSURE™ 0.2 µm • Bioburden reduction applications • Bioburden reduction filter
(PES) PNA Series Single Layer • In-process filtration • Membrane is rated to >7 LRV for
cross-contamination risk from in-process liquids such as buffers. Cell Filters B. diminuta
culture growth media are filtered using 0.1 μm filters to mitigate the risk • 100% integrity tested prior to
of mycoplasma contamination. Since biopharmaceutical unit operations release
are discontinuous in nature, elution pools (e.g. Protein A pool) need Nylon 66 LifeASSURE™ 0.2 µm • Retention of negatively charged • Sterilizing Grade Filter
SP Series Filters Charged particulate contaminant including • Rated to remove 107 B. diminuta
to be filtered with 0.2 μm filters prior to storage in order to manage
Modified endotoxins per ASTM ASTM F838-05 Tests
bioburden loads. To prevent growth of microorganisms in downstream • Suitable for removal of endotoxins in • 100% integrity tested prior to
chromatography steps, post depth filter permeate is filtered using water at point of use release
0.2 μm sterilizing or bioburden reduction grade filters. Performance Nylon 66 LifeASSURE™ 0.1 µm • Rated to provide LRV 7 of • Validated 0.1 µm membrane
PSA Series mycoplasma while providing good that provides reliable retention of
characteristics of LifeASSURE™ PDA PES membranes in various Figure 13: Biopharmaceutical — Sterile Final Filtration and Filling Process Filters flow rate mycoplasma
biopharmaceutical fluids are shown in graphs 6, 7 and 8. • 100% integrity tested prior to
release
PTFE LifeASSURE™ 0.2 µm • Sterile air venting and filtering • Sterilizing Grade Filter
PFS Series applications • Rated to remove 107 B. diminuta
3M Sterilizing Grade 0.2 µm Membrane Filters: Filters • Liquid validation of B. diminuta per ASTM ASTM F838-05 Tests
retention to provide reliable sterilizing • 100% integrity tested prior to
• LifeASSURE™ PSA—Nylon performance in wet or dry conditions release
• Demonstrated complete aerosol
• LifeASSURE™ PFS—PTFE 50 mm Capsule Filters For Small-Scale Sterilizing Grade Liquid Filtration retention of the bacteriophage
Φ X-174
™
• LifeASSURE PDA—PES
Graph 6: Water Flow Graph 7: Bovine IgG Graph 8: CHO Cell Media

Table 6 Comparative Choice Of Membranes†

Nylon PES PTFE
• Good wettability • High flow rates • Hydrophobic
• Positive charge • Great throughput • Solvent stable
Strengths • Lower rates of false integrity test • Low and high pH compatibility
(IT) failure • Gamma stable
• Low protein adsorption
• Lower flow rates • Wettability • Difficulties conducting integrity
• Low pH compatibility • Need to autoclave / steam wet tests, such as the need to wet the
Weakness • Nonspecific adsorption membrane with alcohol
• Gamma tolerance Relative water flow of 10 inch LifeASSURE™ Relative throughput of bovine IgG (50 mg/ml) Relative throughput of CHO cell media feed with
PDA and commercially available PES cartridge with equal area polyethersulfone membrane. equal area polyethersulfone membrane. Tests were
• Many cytotoxic drugs, • Biotech products • Critical vent filtration filters. PES A, B, C cartridges water flow from Relative filtrate volume after 10 minutes at 15
Lead applications • LVP applications • Buffer / media filtration • Solvent / Sterile API applications
conducted at a constant flow and throughput volume
product literature. psi feed pressure was measured. was measured at a terminal pressure of 25 psid.

18 19
PRAXSA

3M makes innovative products for biopharmaceutical monitoring applications. 3M Comply™ chemical indicators can provide
a visual ‘Accept’ / ‘Reject’ for steam sterilization. Our innovative Attest™ Rapid readout biological indicators can provide rapid
results when using biological indicators while simultaneously avoiding aseptic transfers, and the need to prepare media. Clean-
Trace™ technologies use ATP monitoring to test for biological activity on surfaces and to validated the cleaning process. 3M 3M™ Clean-Trace™ Technologies
Pertrifilm Aqua Plates are suitable for efficient monitoring of plant’s water.  ATP indicates biological residues — food, bacteria, body
fluids
Why wait 24 hours for biological results?  ATP tests are fast — as little as 30 seconds
The difference between  Easy to use hardware and software
3M™ Attest™ 1292-S Rapid Readout Biological Indicators and Conventional BI Technology and Rapid Readout BI Technology

3M™ Attest™ Auto-readers Conventional 3M™ Attest™ Rapid Readout Proven Real-Time Results You Can Rely On To Make
Biological Indicators BI Technology Critical Business Decisions
 Provide final readout in 3 hours to validate steam sterilization applications Bacterial enzymes break down Spore-associated enzyme breaks down
 Consistent and repeatable results with high levels of sensi-
nutrients in media... nutrients attached to a fluorescent dye.
 Self-contained biological indicator design reduces risk of contamination during During breakdown of nutrients, tivity demonstrated in independent studies
transfer
the fluorescence 1 Hour
24
is released. 1291
Blue Cap  Used around the world by leading food and beverage manu-
 Recognized by the US Food and Drug Administration as a biological indicator to facturers as an integral part of their HACCP plans
48 3 Hour
 Auto-reader has either 12 or 36 vial capacity and provides automatic calibration on Hours
1292
Brown Cap
 Can be tailored to suit requirements of large manufacturing
Nutrients allow
every biological indicator microorganism to 1 to 4 hours later. . . facilities or smaller laboratories
grow and multiply.
4 Hour
1294
Green Cap Increased Operational Efficiencies
 Powerful data trending software supplied with the 3M™
Clean-Trace™ NG Luminometer instrucment to easily track
and improve hygiene performance over time
Microorganisms give
off acid waste. Fluorescence is detected by Auto-reader.  Provides information to optimize cleaning regimes which
24 to 48 hours later . . .
can generate savings in cleaning materials and labor requirements
When enough acid accumulates
pH-sensitive dye turns yellow.
Same-day results can be up to  Helps create production environment conditions conducive for maximizing shelf life of
95% faster than conventional BIs! products

Chemical Integrators For Monitoring Steam Sterilization 3M™ Petrifilm™ Aqua Plates for Water Testing
 “Accept” or “Reject” at a glance 3M™ Petrifilm™ Aqua Plates are sample-ready media that replace Advantages:
 For use in all 118-138°C (245-280°F ) steam sterilization cycles conventional agar, petri dishes, media pads and disposable filter
 80% increase in productivity
funnels used in the microbial testing of water. Each plate contains a
 Conform to ANSI/AAMI/ISO 11140-1:2005 and EN ISO 11140-1:2005 Class 5 Integrating • Ease of use
water-soluble gelling agent, nutrients and indicators in a dry, shelf-
Indicators • Compatible with membrane filtration
stable format.
 Identify non-sterile instruments before they enter the sterile field  85% savings in storage space
We offer four 3M Petrifilm Aqua Plates to cover your
unique testing needs: (50 3M Petrifilm Aqua Plates vs.
50 agar dishes and 50 disposable filter funnels)
 Heterotrophic Count
 Coliform Count  Confirmed coliform results in just 24 hours

 Enterobacteriacea Count  Longer shelf life vs premade agar plates

 Yeast and Mold Count

20 21
www.praxsasrl.com info@praxsasrl.com
Scientific Applications Support Services (SASS)
3M has a global team of market-focused scientists and engineers who excel in supporting, collaborative efforts between our customers
and 3M. Our technical team is skilled in performing on-site bench-scale tests and relating their results to full scale manufacturing filter
operations. When unique processing problems are encountered, our expert product and application specialists are equipped to identify
filter solutions using either 3M’s broad array of existing products or work directly with our customers to design a custom solution for
the job.

Te
s tin
gS
er ss Validation Lab S
vic t Proce ervi
es ces
es ervic tC
Su

y S om
ol g m
pp

er
no
or

3M Purification provides

cia
ch
tS

Te

lt
comprehensive technical services
er

Fie
and advanced engineering support

w3
vic

ld T
China

for biopharmaceutical customers

/ Ne
es

echn
from centers around the world.

t QA t Strategic
Switzerland

ical Serv
Meriden, CT Japan

France India

St. Paul, MN
Singapore

ices/Ap
Produc

p
Poland

li
Australia

cations
t
Brazil

e
Validation Services

ic

t
r v
3M provides the following validation and scientific services for the biopharmaceutical industry from its various regional global

Pro
Se
technical service centers.

d
er

u
m

ct
Major Technical Centers

En
st
Post-use Integrity Test Compatibility Extractables Cu
gin
ee
GMP Guidelines Sterilization Flushing rt t rin
o
Process Fluid CFR Biological Safety pp gt
Safety Margin USP USP
Sec
ond L ces/Su
evel Technical Servi
Correlation Integrity Toxicity
Hardware Identity
Bacterial Challenge Test Permitted materials End Notes
ASTM Adsorption GMP Guidelines 1 King, David. Applications And Engineering Of Monoclonal Antibodies. Philadelphia, PA: Tyler & Franics Inc., 1998. Print.
B. Dimunuta ATCC 19046 GMP Guidelines Flushing 2 Yigzaw, Yinges; Piper, Robert; Tran, Minh; and Shukla, Abhinav A. “Exploitation of the Adsorptive Properties of Depth Filters for Host Cell Protein Removal
CFR during Monoclonal Antibody Purification.” Biotechnology Progress. 2006: 22, 288-296. Web.
HIMA Binding 3 Robinson, Andrew; Hudson, Michael; and Carnage, Martin. Vaccine Protocols. Totowa, NJ: Humana Press Inc. 2003. Print.
Procedures Preservatives Quantity 4 Dübel, Stefan (Editor). Handbook of Therapeutic Antibodies, Volume 1. Germany: Technical University of Braunschwieg Institute of Biochemistry and
GMP Guidelines Equilibrium Active Materials Biotechnology. 2007. Print
Process Operating Conditions Saturation 5 Dorin, Glenn J. et al. (2001) Method of solubilizing, purifying, and refolding protein. US Patent 6,319,896. Page 46-47.
6 Lin, Jing. “Lipid Foulant Interactions During Chromatographic Purification of Virus Like Particles (VLP) from Saccharomyces Cervacie.” Ph D Thesis, University
Worst case Recovery College London, Sept 2010.
Minimum Qualifying Standards † The information presented here contains data and conclusions from studies where process conditions of filtration and separation technology was optimized by the
Grouping / Bracketing respective researchers. Each application of filtration and separation technology may have significant process differences and it is important for the user to conduct
Correlated Integirty Tests similar process and scale-up studies to validate performance in a particular application. Please contact 3M Purification's global SASS team for help with specific
Filter manufacter's initial qualification applications.

22 23
 PRAXSA

Important Notice
The information described in this literature is accurate to the best of our knowledge. A variety of factors, however, can affect the performance of the Product(s) in a particular
application, some of which are uniquely within your knowledge and control. INFORMATION IS SUPPLIED UPON THE CONDITION THAT THE PERSONS RECEIVING THE SAME
WILL MAKE THEIR OWN DETERMINATION AS TO ITS SUITABILITY FOR THEIR USE. IN NO EVENT WILL 3M PURIFICATION INC. BE RESPONSIBLE FOR DAMAGES OF ANY
NATURE WHATSOEVER RESULTING FROM THE USE OF OR RELIANCE UPON INFORMATION.
It is your responsibility to determine if additional testing or information is required and if this product is fit for a particular purpose and suitable in your specific application.
3M PURIFICATION INC. MAKES NO REPRESENTATIONS OR WARRANTIES, EITHER EXPRESS OR IMPLIED INCLUDING WITHOUT LIMITATION ANY WARRANTIES OF
MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE OR OF ANY OTHER NATURE HEREUNDER WITH RESPECT TO INFORMATION OR THE PRODUCT TO WHICH
INFORMATION REFERS.

Limitation of Liability
3M Purification Inc. will not be liable for any loss or damage arising from the use of the Product(s), whether direct, indirect, special, incidental, or consequential, regardless of the
legal theory asserted, including warranty, contract, negligence or strict liability. Some states do not allow the exclusion or limitation of incidental or consequential damages, so the
above limitation may not apply to you.

Your Local Distributor:

3M Purification Inc. 3M, Attest, CleanTrace, Comply and Petrifilm are

400 Research Parkway trademarks of 3M Company.
Meriden, CT 06450, U.S.A. LifeASSURE, MiniCheck and Zeta Plus are
Tel (800) 243-6894 trademarks of 3M Company used under license.
(203) 237-5541 © 3M 2013. All rights reserved.
Fax (203) 630-4530 70-0201-8680-8
www.3Mpurification.com REV 0113

www.praxsasrl.com info@praxsasrl.com