Bioprocessing & Microbial Technology
Bioprocessing & Microbial Technology
Bioprocessing & Microbial Technology
Teaching
hours/week
Sl.
No.
Duration
L
T
P
(Hrs.)
Theory Subjects
Examination
End
C.I.E.
Exam
Marks Marks
Credits
Total
Marks
FBE011
Upstream Process
---
---
40
50
50
100
FBE012
Biopharmaceuticals and
Facility Engineering
---
---
40
50
50
100
FBE013
---
---
40
50
50
100
FBE014
40
50
50
100
FBEL011
--- --Laboratory
--8
160
50
50
100
FBEL012
---
160
50
50
100
Regulatory Affairs
Bioseparations
Techniques/Downstream
Upstream Laboratory
Downstream processing
laboratory
Upstream Process
Contact Hours /Week
Total Lecture Hours
Total Tutorial Hours
Sub. Code
: 4 Hrs.
:40
:0
:0
:FBE011
Credits
CIE Marks
SEE Marks
:4
: 50
: 50
Module I 7 hrs
Introduction to bioprocess & fermentation technology: Interaction between chemical
engineering, Microbiology and Biochemistry. History of fermentation. Introduction to
fermentation processes, Microbial culture, Screening and selection for fermentation
processes; Preservation and improvement of industrially important microorganisms,
Inoculum production for bacterial and fungal processes. Strain development by
mutagenesis, protoplast fusion and transformation of cloned genes.
Module II - 7 hrs
Raw material and media formulation for fermentation process: Fermentation mediaNatural media, synthetic media sources of Carbon, Nitrogen and vitamins, formulations,
antifoams and optimization. Microbial growth kinetics: Microbial growth cycle,
measurement of growth, Batch culture, continuous culture, fed-batch culture, applications
and examples, scale up of fermentation processes. Sterilization of media, Solid Substrate
Fermentation; Process parameters and measurement techniques: measurement of
temperature, pressure and pH, DO, foam etc.; flow rate of liquid and gases; Automation
(processes computerization).
Module III - 8 hrs
Fermentors: Design of a Fermentor: Functions, construction, and maintenance of aseptic
conditions. Control of various parameters: temperature control; Aeration and agitation
system (Non-newdonian fermentations), baffles; types of fermentors, computer
applications in fermentation technology. Sterilization of fermentor, aseptic inoculation
and sampling methods, Specialized bioreactors: tubular bioreactors, membrane
bioreactors, tower bioreactors, fluidized bed bioreactors, Immobilized system and packed
bed reactors and Photobioreactor.
Module IV - 9 hrs
Microbial fermentation: Industrial fermentation, fermentation growth kinetics,
sterilization of Fermentor and validation issues, media for industrial fermentation,
inoculum development for industrial fermentation, aseptic operation of industrial
Fermentor, validation of Fermentor, fermentation process control for the production of
commercially important compounds, aeration and agitation, maintenance and calibration
of sensors used in fermentation, scale up of fermentation process, scale down methods for
2
: 4 Hrs.
: 40
:0
:0
:FBE012
Credits
CIE Marks
SEE Marks
:4
: 50
: 50
Module I 9 hrs
Biopharmaceuticals: Introduction to Biopharmaceuticals and pharmaceutical
biotechnology, History of the pharmaceutical industry, the age of biopharmaceuticals,
Biopharmaceuticals: current status and future prospects, Traditional pharmaceuticals of
biological origin - Pharmaceuticals of animal origin, Pharmaceutical substances of plant
origin, and Pharmaceutical substances of microbial origin.
Biosimilars - Introduction, current status and future (Growth hormones, Interferon, MAbs,
EPO, Insulin, Interleukins, Colony Stimulating factors etc.); Fermentation process
development for Small molecules primary and secondary Metabolites (Organic acids,
solvents, antibiotics, enzymes, polysaccharides, lipids, Pigments and aroma, steroids;
subtilisin,chymosin,vitB12,hepatitis B vaccine, erythropoietin.
Sources of biopharmaceuticals: E. coli as a source of recombinant, therapeutic proteins,
Additional production systems: yeasts, fungal production systems, Host-Vector
Interactions in Escherichia coli, Parameters Influencing the Productivity of Recombinant
E. coli Cultivations. Production of pharmaceuticals by genetically engineered cells
(hormones, interferon). Microbial transformation for production of important
pharmaceuticals (steroids and semi-synthetic antibiotics). Techniques for development of
new generation antibiotics.
Module II - 8 hrs
Cleanroom - What is a Cleanroom? , The Need for Cleanrooms, Types of Cleanrooms,
What is Cleanroom Technology? The History of Cleanrooms.
Basis of Cleanroom Standards, Federal Standard 209, Pharmaceutical Cleanroom
Classification - EU Guide to Good Manufacturing Practice, Guideline on Sterile drug
products.
Information Sources - The International Confederation of contamination Control
Societies (ICCCS) International Cleanroom Standards- IS0 standards, Federal Standard
209E, Pharmaceutical standards. Recommended Practices and Guides of the Institute of
Environmental Sciences and Technology (IEST), Sources of Pharmaceutical Cleanroom
Documents, International Cleanroom Forum.
pre-change zone, Pre-change zone, Changing zone, Cleanroom entrance zone, Exit
Changing Procedures
Cleanroom Clothing Sources and Routes if Inert Particle Dispersion, Routes and
Sources of Microbial Dispersion, Types of Cleanroom clothing, Cleanroom Masks and
Gloves, Cleaning a Cleanroom
References:
1. Clean Room Technology Fundamentals of Design, Testing and Operation by W
Whyte, John Wiley & Sons, 2001.
2. Bioprocess Engineering: Systems, Equipments and Facilities by Linderson,
DElia, Nelson, John Wiley 1994.
3. Bioprocess Engineering: Basic Concepts, 2nd Edition by Michael L.Shuler and
Fikret Cargy, Prentice Hall, 2002.
4. Bioprocess Engineering Principles by Pauline M Doran. Academic Press, 1995.
5. Fermentation and Biochemical Engineering Handbook Principles, Process
Design and Equipments, Henry C. Vogel and Celeste L. Todaro, Noyes
Publication, 1997.
Regulatory Affairs
Contact Hours /Week
Total Lecture Hours
Total Tutorial Hours
Sub. Code
: 4 Hrs.
:40
:0
:0
:FBE013
Credits
CIE Marks
SEE Marks
:4
: 50
: 50
Module I - 7 hrs
Quality life cycle- Introduction; Research and development (Pre-clinical studies, Clinical
studies); Good laboratory practice (GLP) -GLP in Europe, GLP in the UK, GLP in the
USA; Good clinical practice (GCP) - GCP in the USA, GCP in Europe, ICH guidelines
on GCP, Manufacture of clinical trials material; Drug registration in the European Union
Drug registration in the USA; Unlicensed medicines; Good manufacturing practice
(GMP); Good distribution practice (GDP).
Module II 8 hrs
Quality assurance and control Introduction; Relationship between quality
management, QA, GMP and QC; Definition of quality management; Definition of quality
assurance; Definition of quality control; Responsibilities of QA QA requirements in EU,
PIC/S, WHO and FDA; Responsibilities of QC.
Quality systems: ISO 9000 series; ISO 14000 series; Use of the quality systems approach
within the pharmaceutical industry (Quality systems inspection technique (QSIT), Drug
manufacturing inspections.
Good manufacturing practice - Definition of GMP; Different versions of GMP (UK,
European Union, USA, Australia , WHO ,Arab World ); Comparison of different GMP
guidelines; Responsibilities under GMP; Rules versus guidelines; Application of GMP
(Quality management, Personnel, Premises and equipment, Documentation , Production,
Contract manufacture and analysis, Complaints and recalls, Self-inspection).
Good distribution practice - Principles of GDP; Quality system; Personnel;
Documentation; Premises and equipment;
Deliveries to customers; Returns; Self-inspections; Provision of information to Member
States; The role of the responsible person (Statutory basis, Eligibility and duties of the
responsible person, The responsible person and the management representative).
Module IV - 8 hrs
Pharmaceutical microbiology - Overview of micro-organisms ; The microbiology of raw
materials; Non-sterile products; Sterile products ( Small volume potentials, Large volume
potentials, Ointments and creams, Eye drops, Medical devices); Sterile
Manufacturing(Area classification, Terminally sterilized products, Aseptic manufacturing,
Mixed manufacturing); Risks of contamination and their control (Personnel, Premises,
Equipment, Operation of the facility, Cleaning and sanitation); Validation and
environmental monitoring (Monitoring of the air, Monitoring of surfaces, Monitoring of
personnel, Media trials); Sterilization ( Sterilization by heat, Irradiation, Ethylene oxide
sterilization, Filtration, Validation and monitoring of sterilization cycles).
Module V- 8hrs
Quality improvement Programmes - Elements of TQM, TQM and the pharmaceutical
industry, Definition; Prerequisites for successful quality improvement Programmes
(Management support, Management commitment, Team working, Asking the right
question, Breaking out of the silos, Acceptance of change, User driven)
Quality tools and techniques - Tools for identification of problems; Tools for the analysis
of problems; Benchmarking; Using the tools and techniques (Type of quality teams,
Team working the problem-solving approach, Team working the process skills).
References:
1. Biopharmaceuticals Biochemistry and Biotechnology, 2nd Edition, by Gary
Walsh, John Wiley, 2003.
2. Pharmaceutical Engineering Series: Quality and Regulatory Compliance by Kate
McCormic, Butterworth and Heinemann, 2002.
3. Biopharmaceuticals, An industrial Perspective by Gary Walsh and Brendan
Murphy, Kluwer Academic Publishers, 1998.
Bioseparations Techniques/Downstream
Contact Hours /Week
Total Lecture Hours
Total Tutorial Hours
Sub. Code
: 4 Hrs.
:40
:0
:0
:FBE014
Credits
CIE Marks
SEE Marks
:4
: 50
: 50
Module I: - 8 hrs
Basic concepts of Bio-separation Technology, Separation characteristics of proteins and
enzymes size, stability, properties; purification methodologies Characteristics of bio
products; Flocculation and conditioning of broth, overview of reaction processes
involved in separation, numerical examples illustrating the process. Mechanical
separation processes; Filtration at constant pressure and at constant rate; empirical
equations for batch and continuous filtration, centrifugal and cross-flow filtration,
Methods for extraction of proteins - Cell distribution methods for intracellular products
Osmotic shock, Homogenization, The Polytron/UItra-Turrax-type homogenizer, under
high pressure, Sonication, Micro-cavitations, Enzyme digestion. Centrifugation: basic
principles, design characteristics; ultracentrifuges; principles and applications.
Module II: - 8 hrs
Membrane based separation processes, Micro-filtration; Reverse osmosis, Nanofiltration,
Ultrafiltration and Affinity ultrafiltration, concentration polarization, rejection, flux
expression, membrane modules, dead-ended and cross-flow mode, material balances and
numerical problems.
Module III: - 8 hrs
Product Resolution/Fractionation - Chromatography: Gel filtration chromatographyeffect of particle size, mobile phase flow rate, hydrostatic pressure, sample preparation,
loading sample, molecular weight determination, spin columns. Equipment required for
low pressure liquid.
Chromatography: Ion-exchange chromatography (IEC), Ion-exchange resins,
Chromatofocusing, Molecular exclusion chromatography (MEC), Determination of MW
by MEC. Hydroxyapatite chromatography,. Affinity chromatography: Immunoaffinity
purification, Immunoaffinity matrices, ligand affinity. Adsorptive chromatographic
separations processes, hydrophobic interaction chromatography (HIC), RP - HPLC
Module IV : - 8hrs
Analysis of the final product - Protein-based contaminants, Removal of altered forms of
the protein of interest from the product stream, Product potency, Determination of protein
concentration, Detection of protein-based product impurities, High-pressure liquid
chromatography (HPLC), Mass spectrometry, Immunological approaches to detection of
contaminants, Amino acid analysis, Peptide mapping, N-terminal sequencing, Analysis of
secondary and tertiary structure, Endotoxin and other pyrogenic contaminants, Endotoxin
- the molecule, Pyrogen detection, DNA, Microbial and viral contaminants, Viral assays,
Miscellaneous contaminants, Validation studies
Module V: - 8 hrs
Final product formulation, Some influences that can alter the biological activity of
proteins, Proteolytic degradation, Protein deamidation, Oxidation and disulphide
exchange, Alteration of glycoprotein glycosylation patterns Stabilizing excipients used in
final product formulations, Final product fill, Freeze-drying, Labeling and packing.
References:
1.
2.
3.
4.
5.
10
: 16Hrs.
: 160
:0
:0
:FBEL011
Credits
CIE Marks
SEE Marks
:3
: 50
: 50
Shake
flask
optimization
11
12
Downstream Laboratory
Contact Hours /Week
Total Lecture Hours
Total Tutorial Hours
Sub. Code
: 16 Hrs.
: 160
:0
:0
:FBEL011
Credits
CIE Marks
SEE Marks
:4
: 50
: 50
Downstream processing
Centrifugation, Filtration: TFF principles and function, Membrane selection criteria,
Diafiltration, Cleaning and sterilization
Chromatography: Affinity Chromatography, Hydrophobic Interaction, Ion Exchange, Gel
Filtration.
Cleaning-in-Place (CIP), Sterilization-in-Place (SIP), Sterilization by Filtration.
Validation - Autoclave, Hot Air Oven; WFI (conductivity, endotoxin testing); swap test
of vessels.
Electrophoretic techniques, Protein quantification techniques
1. Centrifugation 20hrs
Functions and process optimization for centrifugation processes at variable speed and
flow rate, Continuous centrifugation for cell concentration or cell debris removal,
Cleaning and sterilization of continuous centrifuge, Microbial concentration / separation
analytical method by spectrometry and assay validation
2. Cleaning-In-Place 20 hrs
Familiarization with process vessel, CIP processes, Generation of P& ID and valve
matrix for CIP processes, OQ of process vessel: spray ball coverage test with riboflavin
3. Sterilization-In-Place 25 hrs
SIP processes, Generation of P&I D and valve matrix for SIP processes, SIP PV: Heat
penetration study for ESIP and FSIP using temperature probes / biological / chemical
Indicator.
13
techniques
SDS-PAGE
14