1. Chapter 1- Introduction to Biomanufacturing
Northeast Biomanufacturing
Center and Collaborative

2. What is Biomanufacturing ?
Advanced-technology manufacturing industry within biotechnology that is responsible for making biopharmaceuticals (biologics) and other bioproducts such as biofuels and human tissues.
A biopharmaceutical/biologic is any biotechnology-based therapeutic that structurally mimics components found in a living organism.

3. Examples of Biopharmaceuticals
Hormones (Insulin)
growth factors
blood proteins
clotting factors (tPA)
enzymes
antibodies
DNA and RNA
stem cells

4. Applications of Biopharmaceuticals in Health & Medicine
Therapeutic proteins for treatment of disease
Vaccines to prevent disease
Protein or DNA-based diagnostics
Regenerative medicine technologies
Gene therapy

5. Approved Biologics Biopharmaceutical
Date of Commercial Production, Indication
Company
Humulin (human insulin)
1982, diabetes
Eli Lilly
tPA (tissue plasminogen activator
1987, myocardial infarction
Genetech
Humatrope (human growth hormone)
1987, human growth hormone deficiencies
Epogen
1989, anemia
Amgen
Energix-B
1989, hepatitis B vaccine
SmithKline Beecham
Betaseron
1989, multiple sclerosis
Berlex Laboratories/Chiron
Ceredase
1991, Type I Gaucher’s disease
Genzyme
Proleukin (Il-2)
1992, kidney carcinoma
Chiron
AHF (recombinant anti-hemolytic factor)
1992, hemophilia A
Baxter Healthcare
Pulmozyme (DNAase)
1993, cystic fibrosis
Genentech

6. Biopharmaceutical Drug Development
Objective
To develop a robust, scalable, reproducible, and cost-effective process that results in safe and efficacious biopharmaceuticals
Time consuming and expensive!
8-15 years
$500 million to $1 billion
1/10,000 drug candidates makes it

7. Drug Development Process

8. Discovery/ Research Identify gene/protein of interest
Create an “expression vector” that includes the gene of interest
Select cell type to transform
Cellular machinery transcribes DNA into mRNA and mRNA into protein via the Central Dogma

9. Expression Vector: The basis of modern biopharmaceutical manufacturing

10. Bacterial Transformation in Biomanufacturing

11. Central Dogma of Biology

12. Expression Systems- Considerations
Post-translational modification
Modification of an amino acid of the polypeptide chain
Addition of sugar moieties to certain amino acid side chains(glycosylation)
Approximately 70% of human proteins are glycosylated
and biological activity is highly affected without this modification
Majority of prokaryotic cells lack the ability to make
glycoproteins

13. Prokaryotic Expression Systems
Bacterial Systems (e.g E.Coli)
Less time consuming/expensive to culture
Unable to perform post-translational glycosylation
Yeast Cultures
Less time consuming/ Inexpensive
Can add non-human glycans which are immunogenic

14. Recombinant DNA Expression Systems
Application (%)
CHO
35.5
E. Coli 29
Miscellaneous mammalian
16.5
Yeast
Miscellaneous
8.5
Human 4

15. Blockbuster Drugs made in CHO cells
Product
Sales
(USD in billions)
Approved
Patent Expiry (US)
Company
Humira (anti- TNF)
11.0
2002
2016
AbbVie, Eisai
Enbrel (anti-TNF)
8.76
1998
2026
Amgen, Pfizer, Takeda
Rituxan (anti CD20)
7.91
1997
Biogen Idec, Roche
Avastin (anti-VEGF)
6.97
2004
2017
Roche/Genentech
Herceptin (anti-HER2)
6.91
1996
2019
Epogen (epoetin alfa)
3.36
1989
2013
Amgen, Johnson & Johnson
Avonex (IFN-b-la)
3.00
Biogen Idec
Rebif (IFN-b-la)
2.59
Merck Serono
Aranesp/Nesp
2.42
2001
2024
Amgen
Eyla
1.86
2011
2021
Regeneron; Bayer Healthcare
Table taken from website Brief history of CHO cells

16. Eukaryotic Expression Systems
Mammalian Systems – Chinese Hamster Ovary Cells (CHO)
Have the ability to perform post-translational glycosylation
Widely used but expensive and more time consuming to culture
Protein products are easier to process later in production because they are secreted into culture media

17. Process Development and Scale-Up
Determine best processes to produce the most protein/drug as efficiently as possible.
Upstream Processing- develop equipment and processes used in production to culture the cells that make the biopharmaceutical
Downstream Processing –recovery & purification of drug from cell culture media
Formulation/Distribution –product is filled, tested for purity and sterility and distributed for clinical use

18. Upstream Processing Culture Flasks Spinner Flasks Bioreactors
Develop equipment and processes used in production to culture the cells which produces the biopharmaceutical
Culture Flasks
Spinner Flasks
Bioreactors

19. Downstream Processing
Recovery and purification of drug product from cell culture
Centrifugation- Ultracentrifuges
Filtration-TFF Units
Chromatography- FPLC/HPLC

20. Testing Finished Product
The biopharmaceutical is placed in solution, with excipients, for parenteral injection. Product tested for purity, stability, and sterility before distribution
Purity Tests
Chromatography
Electrophoresis
ELISA
Sterility Tests
Microbial Tests for endotoxins

21. Regulation of Biopharmaceutical Manufacturing
The manufacture of all pharmaceutical products are regulated by the Food & Drug Administration (FDA) to ensure quality and safety.
FDA requires that all pharmaceuticals be manufactured and quality of drug products using Current Good Manufacturing Processes or cGMP

22. Ensuring Quality
Drug Manufacturing Facilities have departments/groups that ensure quality:
Quality assurance (QA) Dept ensures that the manufacturing facility complies to all regulations and maintains all documentation for compliance
Quality Control (QC) Dept is responsible for testing the raw materials and the product during many stages of the manufacturing process

23. Documentation in Pharmaceutical Manufacturing
Documentation can take many forms:
Process and production document (SOPs, batch records, manufacturing operations)
Operational and equipment log books (documentation of maintenance, cleaning)
Training documents
Electronic Documents

24. Biopharmaceutical Biomanufacturing
The production of a new biopharmaceutical may require a facility to be prepared for its manufacture and 400 to 600 people hired; 50% may be technicians
The largest bioreactor in such a facility could be 20,000 L (scaled-up is from a 1-2 mL cryovial taken from the working cell bank)
Once constructed and commissioned, the facility’s equipment and process SOPs must undergo validation
The set up, maintenance and use of each piece of equipment is logged; the equipment must be calibrated
Environmental Health and Safety (EH&S) requirements are of central importance.

25. Amgen-Biomanufacturing Facility Tour
Visit Amgen website for a virtual tour of their manufacturing facility

26. Top of 20,000L Bioreactor