Lecture 9a scaling up

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Industrial Microbiology Dr. Butler 2011

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Lecture 9a scaling up

  1. 1. Lecture 9 Animal Cell Biotechnology Scaling up the production processScaling up animal cell/microbial processes• optimal physiological conditions obtained via small scale processes maintained for large scale operation• must consider: 1. fermentor/bioreactor system 2. control of agitation, pH, temperature, dissolved oxygen
  2. 2. Lecture 9 Animal Cell BiotechnologyScaling up the production process T-flasks
  3. 3. Lecture 9 Animal Cell BiotechnologyScaling up the production process Spinner bottle
  4. 4. Lecture 9 Animal Cell Biotechnology Scaling up the production processMultiple process vs. unit process• can have 1000 100 mL flasks or a single 100 litre fermentor (same volume)1. Multiple process• easy to replicate samples and conditions, large surface area/volume ratio, tedious to take care of all samples, labor intensive2. Unit process• good for large scale production of biological products, ease of handling, decreased possibility of contamination
  5. 5. Lecture 9 Animal Cell Biotechnology Scaling up the production processButler, M. 2004. Animal cell culture and technology 2nd ed. London and New York:Garland Science/BIOS Scientific Publishers. P203.
  6. 6. Lecture 9 Animal Cell Biotechnology Scaling up the production processSuspension vs. Anchorage-dependence• suspension cells are readily scaled up to production level using similar fermentation vessels as microbial cells (with modifications)• anchorage-dependent cells require a surface for attachment in order to grow → grow as static cultures (i.e. hollow fibres) → grow on microcarriers, in a stirred tank reactor (fermentor)
  7. 7. Cell culture processes• Anchorage-dependent systems Microcarrier (dextran, collagen or plastic) : 200 µm Advantages: -Higher cell yields - increased productivity. - ↑ surface ratio (100 -200 cells). - suspension cultures• Suspension systems: > Industrial culture
  8. 8. Cytopore microcarriers
  9. 9. Cell Culture system• Vero cells •Why? Regulated by WHO • already licensed for vaccine production such as polio, influenza, rabies, others• Serum-free medium •Why? Safety and reproducibility• Microcarriers •Why? Scalability and ease of infection
  10. 10. Macroporous microcarrier
  11. 11. Lecture 9 Animal Cell Biotechnology Scaling up the production processUsually scale up 10x at a time: 1 → 10 → 100 → 1000 → 10,000 LFactors limiting scale-up• supply of oxygen• shear damage from mixing• build up of toxic metabolites
  12. 12. Lecture 9 Animal Cell Biotechnology Scaling up the production processduring scale up decreased product yields may be experienced due to the following:1. inoculation – must consider quality and quantity of inoculum used to start process $2. choice of medium – cheaper materials often used for large $ $ scale production due to cost limitations - omission / reduction of serum, antibiotics3. large scale sterilization – longer sterilization times may result in degradation of heat labile compounds, reducing quality of media4. development of gradients - larger fermentors/bioreactors may be subject to development of nutrient, temperature, pH, and oxygen gradients5. other factors -scaling up may also alter the generation of foam, shear forces
  13. 13. Lecture 9 Animal Cell Biotechnology Scaling up the production process: The Stirred Tank Reactor• bioreactor is another term for fermentor• stirred tank reactor simplest and most widely used system (pot and paddle) → small vessels (<20 litres) made of glass → large vessels (>20 litres) made of stainless steel
  14. 14. Bioreactor (Fermenter)• Vessel that allows the growth of cells.• Stirred tank bioreactor (STR). Growth: Bacteria, yeast, mammalian cells.• Homogeneous solution mixed by an impeller: -Laminar stirred: Just one dimension to lift the liquid. Cultures >1L: Uneven oxygen exchange and poor nutrient distribution www.corning.com
  15. 15. Stirred Tank Reactor• Shape: -curve bottom mammalian cells -square bottom poor mixing• Materials used: Vessel up to 10 L (glass) Head plate steel stainless• Heat control: (heat pad or jacket water) - Constant To -mammalian cell culture (37o C)
  16. 16. Lecture 9 Animal Cell Biotechnology Scaling up the production process: The Stirred Tank ReactorButler, M. 2004. Animal cell culture and technology 2nd ed. London and New York:Garland Science/BIOS Scientific Publishers. P156.
  17. 17. Fig. 9.2
  18. 18. Fig. 9.3
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