Manufacturing Overview
Biomanufacturing Overview
Biomanufacturing 101
Jim DeKloe
Biomanufacturing Overview 1 Biomanufacturing 101 Welcome to Biomanufacturing 101 Jim DeKloe - Solano College
Jim DeKloe
707-477-8354
james.dekloe@solano. edu
Biomanufacturing Overview Biomanufacturing 101 “Biomanufacturing” versus “Biotechnology”
Biomanufacturing Overview 3 Biomanufacturing 101 Manufacturing means equity
Biomanufacturing occurs in more mature companies
Manufacturing signals equitable participation of members of the community
The participation of “Regular People” as lab technicians and on the manufacturing line
Upward mobility and advancement
Biomanufacturing Overview 4 Biomanufacturing 101 Biomanufacturing is the intersection of
Science
Engineering
Business
Government
Quality
Biomanufacturing Overview 5 Biomanufacturing 101 New Biotechnology
Biomanufacturing Overview 6 Biomanufacturing 101 Biomanufacturing Overview 7 Biomanufacturing 101 Back to Basics - Cells
Biomanufacturing Overview 8 Biomanufacturing 101 Genentech Announces Vacaville Expansion - 1994
Biomanufacturing Overview Biomanufacturing 101 Biomanufacturing
Biomanufacturing Overview Biomanufacturing 101 Biomanufacturing Overview Biomanufacturing 101 Biomanufacturing Overview Biomanufacturing 101 Biomanufacturing Overview Biomanufacturing 101 Unit Operations
All Unit Operations All unit operations must be maintained in must be validated a “state of control”
Biomanufacturing Overview 14 Biomanufacturing 101 Prokaryotic or Eukaryotic? E. coli or CHO cells?
E. coli CHO cells
•Grow fast •Grow slowly
•Use inexpensive medium •Use expensive medium
•Produce High Amounts of Protein •Produce lower amounts of protein
•May not fold properly •Fold proteins properly
•Do not glycosylate •Can glycosylate
•Less contamination risk •More contamination risk
•Less difficult “upstream,”, more •More difficult “upstream,”, less difficult “downstream” difficult “downstream”
Also: yeast, insect, filamentous fungi, human
Biomanufacturing Overview 15 Biomanufacturing 101 Manufacturing Processes
Upstream Testing, Cell Downstream Formulation Cell Labeling, Bank Purification Fill &Finish Culture Pkg
2-3 Weeks 1 Month 1 Week 1-2 Days 1-2 Months + Safety Testing
Quality review of all manufacturing and testing documents, prior to final drug product release for patient use
Biomanufacturing Overview Biomanufacturing 101 Cell Banking Process
Usually a two-tiered system consisting of a Master Cell Bank (MCB) and a Working Cell Bank (WCB) is used to provide the supply of cell substrate for manufacture
Master Cell Bank (MCB Working Cell Bank (WCB) Homogeneous suspension Produced from one vial of the of cells originated from a MCB single clone Intended to expand the Aliquoted in storage vials 1 number of vials for production (e.g. 1 mL each) and stored in a liquid nitrogen freezer Every production lot is started from one WCB vial (identical Intended for the entire life- starting material for cell cycle of the drug culture) Biomanufacturing Overview Biomanufacturing 101 Cell Bank Testing
Morphology Identity Isoenzyme analysis Expression construct (restriction analysis, copy number, integration site, sequencing)
Purity Absence of microbial or viral contaminants (animal-derived media)
Consistent production of the intended product over the cell culture period, End of Stability Production Cells (EPCs) Storage stability of cell banks Genetic stability (Analysis of the expression Biomanufacturing Overview construct in EPCs) Biomanufacturing 101 Cell Culture (Upstream) Process
Start of cultivation with cells Propagation of cells in from WCB (1 mL vial) shake flasks/roller bottles
Media Feeds
Cell separation by Cell-free Cell expansion in filtration and/or harvest bioreactors Production centrifugation bioreactor Biological Physical Chemical Sterility Cell density Temperature Oxygen concentration Bacteria
Cell viability Agitation speed pH Viruses Feed Additions Glucose Mycoplasma Biomanufacturing Overview Biomanufacturing 101 Fermentor versus Bioreactor
There is no fermentation in biomanufacturing
All cells should use cellular respiration to make their ATP – they should never be grown anaerobically
The single most important design consideration for a bioreactor is oxygen delivery
“Fermentation” is a slang term or a historical term – it is commonly used, but it is technically incorrect
Biomanufacturing Overview Biomanufacturing 101 Purification (Downstream)
Removal of product and process related impurities
Elution
Membrane Filtration Cell-free Protein A Virus Inactivation 0.2 µm harvest Affinity & pH Adjustment Chromatography
Elution
Ion Exchange Membrane Adsorber Conditioning Dilution Chromatography Anion Exchange
0.2 µm Filtration Drug Substance is Nanofiltration frozen for Virus Removal Ultra-/Diafiltration Filling long-term Biomanufacturing Overview Concentration/Buffer into bags or storage Biomanufacturing 101 exchange bottles Formulation, Finish & Fill
The purified form of the protein drug is usually "formulated" with carefully selected excipients. The operations that occur subsequent to protein purification, such as freezing of the purified protein bulk, thawing of the bulk, formulation (excipient addition), sterile filtration, filling, freeze-drying, and inspection are commonly referred as "formulation and fill-finish operations".
Protein stability, route of administration and ease of use considerations Excipient additions: ionic strength, buffers, pH, surfactants, sugars, salts, antioxidants, and amino acids Protein concentration: IV versus subcutaneous delivery Liquid vs freeze dried formulation Vial vs prefilled syringe or other delivery systems
Biomanufacturing Overview Biomanufacturing 101 Sterile Fill & Freeze Drying
The formulated bulk is sterile-filtered, filled into product vials, and freeze-dried for stability and storage purposes. The vials are then capped and prepared for inspection.
Sterile Vial Freeze-Dryer Final Filtration of Filling Lyophilization cycle Container formulated bulk capped
Biomanufacturing Overview Biomanufacturing 101 Testing, Labeling and Packaging
In the final stage of the process, the medicine is thoroughly tested to ensure product quality. This also includes a visual/automated inspection of every filled DP container and labeling and packaging.
Testing ensures that the DP meets predefined criteria for Safety, Identity, Strength, Quality and Purity Safety: sterility, endotoxin Strength: protein concentration Purity: DNA, HCP Identity: tryptic (and other) digest, CD, Mass spectroscopy Quality: activity assay, oxidation, glycan profile
Biomanufacturing Overview Biomanufacturing 101 Regulation
Biomanufacturing Overview Biomanufacturing 101 Contamination versus Impurities
Contamination – unexpected
Impurities – expected
. Product related Impurities - variants that differ from the desired product . Process Related Impurities - materials added to the process and cellular components
Biomanufacturing Overview Biomanufacturing 101 Quality Assurance and Quality Control
The law requires a QA/QC department that is separate from manufacturing
They will test:
Raw Materials
In-Process Samples
Utilities
Environmental Monitoring
The final protein product
Biomanufacturing Overview 27 Biomanufacturing 101 The Quality Department should operate independently from manufacturing
Quality by Design
CQAs
CPPs
PATs
DoE
Validation
Pharmaceutical Quality Systems – QA/ QC / Validation
Biomanufacturing Overview 28 Biomanufacturing 101 What do GMPs cover?
Subpart Description A General Provisions
B Organization and Personnel C Buildings and Facilities
D Equipment
E Control of Components and Drug Product Containers and Closures
F Production and Process Controls
G Packaging and Labeling Controls
H Holding and Distribution
I Laboratory Controls
J Records and Reports
K Returned and Salvaged Drug Products
Biomanufacturing Overview 29 Biomanufacturing 101 •More PAT (Process Analytical Technology) Trends in Biomanufacturing •Generation of Cell Lines that secrete •Growth in the biomanufacturing field •Fewer impurities into the DS process
•Process Intensification •Regional product setup – regional supply chains
•Higher cell densities •Continuous processing rather than batch processing
•Higher protein titers = 5 – 10 g/liter •The use of ATF - Alternating Tangential Filtration – for protein purification •Smaller fermentor volumes (enabled by the higher protein titers) •The manufacture of biosimilars
•Increased use of perfusion culture – but shorter perfusions •Application of lean manufacturing principles
•Disposable, single use technologies •Application of CRISPR technology to genetically engineer cells •Closed systems used in both upstream and downstream processes •Streamlined clinical approval strategies
•Removal of animal products (especially serum) from the •Streamlined construction schedules for new facilities manufacturing stream •A dramatic increase in new therapeutic products, cell- •Reduced segregation of different protein products based therapies and gene therapies as examples produced in a plant •Greater emphasis on Contract Manufacturing •Greater simplicity of manufacturing plant design Organizations
•Modular plants and “ball-room” design (where different •Personalized medicines and individualized medicine processes are not physically separated) – this trend is (precision medicine) enabled by closed systems •Increased speed to market •Multiple Processing Steps integrated into a single unit operation •Increased pressure to reduce the cost of medicine
•Reduced capital costs, higher production yields •Manufacturing in and for emerging markets •Increased flexibility Biomanufacturing Overview 30 Biomanufacturing 101