Lecture 9a   scaling up
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Lecture 9a scaling up

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Industrial Microbiology Dr. Butler 2011

Industrial Microbiology Dr. Butler 2011

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Lecture 9a   scaling up Lecture 9a scaling up Presentation Transcript

  • 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
  • Lecture 9 Animal Cell BiotechnologyScaling up the production process T-flasks
  • Lecture 9 Animal Cell BiotechnologyScaling up the production process Spinner bottle
  • 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
  • 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.
  • 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)
  • 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
  • Cytopore microcarriers
  • 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
  • Macroporous microcarrier
  • 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
  • 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
  • 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
  • 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
  • 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)
  • 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.
  • Fig. 9.2
  • Fig. 9.3