Introduction to Bioprocessing Monday, January 19, 8:30 AM-5:30 PM – Tuesday, January 20, 8:30 AM-12:30 PM View sample slides CHI’s Introduction to Bioprocessing training seminar offers a comprehensive survey of the steps needed to produce today’s complex biopharmaceuticals, from early development through commercial. The seminar begins with a brief introduction to biologic drugs and the aspects of protein science that drive the intricate progression of analytical and process steps that follows. We then step through the stages of bioprocessing, beginning with the development of cell lines and ending at the packaging of a finished drug product. The seminar also explores emerging process technologies, facility design considerations and the regulatory and quality standards that govern our industry throughout development. The important roles played by the analytical and formulation in developing and gaining approval for a biopharmaceutical are also examined. This 1.5-day class is directed to attendees working in any aspect of industry, including scientific, technical, business, marketing or support functions, who would benefit from a detailed overview of this field. For full details visit chi-peptalk.com/peptalk_content.aspx?id=140088&libID=140059

1. Introduction to Bioprocessing
Cambridge Healthtech Institute Peptalk
Palm Springs, CA
Presented by Susan Dana Jones and Sheila Magil
BioProcess Technology Consultants
www.bptc.com

2. BioProcess Technology Consultants
 World leader in providing strategic, technical, regulatory, and business consulting
services to biopharmaceutical industry
 Founded in 1994; 20+ year proven track record
 Team of 13 consultants has provided consulting services to over 500 clients across six
continents
Process
Development
Quality &
Regulatory
Manufacturing
Strategy Supply Chain Program
Management Due Diligence
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3. Instructors
 Sheila Magil, Ph.D.
Senior Consultant, BPTC
Sheila has over 20 years of experience in
quality and analytical method development
for biologics, peptides and small molecules.
Her expertise includes quality assurance,
protein and peptide biochemistry, and
analytical development. Sheila has
implemented quality systems and has
managed external analytical and QC
activities for multiple biopharmaceutical
products.
3
 Susan Dana Jones, Ph.D.
Vice President and Senior Consultant, BPTC
Susan is a seasoned biotechnology
entrepreneur with experience in product
development, outsourcing, and strategic
planning. She is a subject matter expert in
cell line development and characterization
for biosimilar, new biopharmaceutical, and
vaccine development programs. She has
broad knowledge of regulatory
requirements for manufacturing products
for human use and has prepared CMC
sections of multiple regulatory
submissions.
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4. Introduction to Bioprocessing Course Content
 Definition of biopharmaceutical products
 Regulatory considerations in bioprocessing
 General timelines for biopharmaceutical product development
 Critical CMC activities:
• Project management and tracking
• Analytical methods
• Expression systems
• Downstream processing and viral clearance/inactivation
• Formulation
• Stability
 Cost considerations
 Process qualification
 Comparability
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5. What are Biopharmaceutical Products?
 Therapeutic proteins including antibodies, nucleic acids, gene therapy viral vectors
• Generally produced by recombinant DNA technology
 Excluded from definition are traditional biologic products
• Non‐recombinant vaccines, plasma‐derived proteins
 Focus of this training course is on production of biopharmaceutical products,
especially monoclonal antibodies, in mammalian cell culture
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Example
Biopharmaceutical Product MW # of Amino
Acids
# of Protein
Chains Glycosylated
Insulin ~6 kDa 51 2 No
Growth Hormone ~22 kDa 191 1 No
Erythropoietin 30‐34 kDa 165 1 Yes
Coagulation Factor IX ~52 kDa 461 1 Yes
Monoclonal antibody (IgG1) ~150 kDa ~660 4 Yes
Factor VIII ~240 kDa 2133 1 Yes

6. Project Management: Integration of Development Activities
When to scale up, what
to use for GLP tox, what
scale, batch records
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Analytical methods:
Understand the
molecule Microbial or
mammalian, novel or
established,
accelerated or
standard?
Expression
System
Platform or de novo,
small scale systems, QbD,
DOE, applied knowledge
Process
Development
Scale up and
Production
Analytical
Methods

7. Protein Structure
 Proteins have an inherent degree of structural heterogeneity
 Primary Structure
• Amino acid sequence
• Held together by peptide (amide) bonds
 Secondary Structure
• Held in place by hydrogen bonding
 Tertiary Structure
• Driven by hydrophobic and ionic interactions
 Quaternary Structure
• Cluster of similar or different proteins
Primary Structure
Secondary
Structure
Tertiary
Structure
Quaternary
Structure
Amino
Acids
‐Pleated
Sheet
‐Helix
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8. Classes of Analytical Methods
Classes of Methods Specific Methods
Spectroscopy UV‐Vis, CD, FTIR, Fluorescence, Raman
Chromatography Reverse Phase, Size Exclusion , Ion Exchange,
Hydrophobic Interaction, Affinity
Electrophoretic SDS‐PAGE and Normal Phase, Reduced and Non‐reduced,
2‐Dimensional, Isoelectric focusing,
capillary Coomassie and Silver stains
Immunophoretic Western Blot, ELISA
Immunoassays Bioassays, ELISA
Potency Bioassays, Cell‐based, Binding
Microbiological Analyses Bioburden, Sterility, Viability, Microbial
Contamination, Endotoxin
Chemical (Compendial) USP/EP/BP/JP raw material testing
Other Analytical Ultracentrifugation, Particulate Matter,
pH, Visual appearance
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9. Role of Analytics
With each step, analytical methods have become increasingly important and front‐end
loaded
“The QbD process design starts with an intensive
characterization of the product through a large array of
biochemical and biophysical analyses at normal and stressed
conditions…”
The QbD process design starts with an intensive characterization of the product
through a large array of biochemical and biophysical analyses at normal and stressed
conditions…”
Banerjee A. BioPharm Intl 23(5): 26‐40.
9 Banerjee AF. rBoiomPha rCmloInntl e23 (t5o): 2C6l‐4in0.ic®
Process defines
Product
Well
Characterized
Biologics
Quality by
Design and
Process
Analytical
Technologies
Increasing process and product
understanding
1980’s 2010’s

10. General Scheme for Biopharmaceutical Manufacturing
Working Cell Bank (WCB)
Inoculum Preparation
Production Bioreactor
Primary Recovery
Purification
Polishing
Formulation
Bulk Drug Substance (API)
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11. Common Production Hosts
Critical issues to address
 Technical
• Overproduction may cause
aggregation or degradation by host,
toxicity to host cell, inaccurate or
incomplete processing
• Post translational modification
(glycosylation)
 Intellectual Property Rights
 Manufacturing Capabilities for Clinical
and Commercial Supplies
 Bacteria
• E. coli
• Pseudomonas fluorescens
 Yeast
• Saccharomyces cerevisiae
• Pichia pastoris
 Mammalian cells
• Chinese hamster ovary (CHO)
• BHK and HEK (specific indications)
• Per.C6 and other human lines
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12. An important trend – single use bioreactors
Xcellerex
XDRTM Bioreactor
Sartorius Stedim
Biostat® Culti‐bag
Thermo Fisher (Hyclone)
Single‐use Bioreactor
ATMI
NucleoTM Bioreactor
GE Healthcare
Wave Bioreactor
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13. Downstream processing of biopharmaceuticals
Downstream Process Bulk
Viruses
Pyrogens
Cells
Nucleic acids
Viruses
Cell
culture
medium
Host cell
proteins
CIP
Nucleic
acids
Leakage
Protein Raw
Material
(rDNA)
Pure Bulk
Product
(Therapeutic)
Ligand
Leakage
Microorganisms
Pyrogens
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14. Column sizing and optimization
 Balance cost of production (COP), production rate (g/hr)
and productivity (g/hr/L support)
• Trade‐off between capacity, throughput, and cost
• Multiple cycles vs. a larger column?
• How frequently should media be replaced?
• Support equipment sizing and cost
Cycling small
column in series
Multiple small
columns in parallel Single large column
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15. Product Comparability
 Primary structure
• Amino acid sequence
 Secondary structure
• Three‐dimensional structure of the
protein (alpha helices, beta sheets,
loops/turns)
 Tertiary structure
• 3‐D structure of protein through
interaction of the secondary
structures
• Increased regulatory emphasis on
this aspect of structure
 Quaternary structure
• Describes the three‐dimensional
arrangement of protein subunits
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16. Product Consistency
Potency
Structure
Stability
Safety
Efficacy
Product Control Through Process Control
Cell line
Media and Feeds
Culture Time
Column Loading
Intermediate Hold
Time
Formulation Buffer
QbD
PAT
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17. Thank You!
Susan Dana Jones and Sheila Magil
BioProcess Technology Consultants, Inc.
12 Gill Street, Suite 5450
Woburn, MA 01801