Term
| What defines a reproducible culture for biomanufacturing? |
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Definition
1. purity
2. high viability (>95% viable)
3. consistent umax under defined growth conitions (predicts length of process)
4. consistent cell yield on a growth limiting substrate (predicts final yield)
5.consistent Ks on a growth limiting substrate (determines when culture is nutrient limited)
6. consistent morphology (determines whether the medium was correctly formulated) |
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Term
What are 6 basic steps in medium optimization using complex nutrients?
What 2 factors are important when optimizing? |
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Definition
1. establish microbes nutritional requirements for growth product formation
2. substrate uptake/transport
3. product/process compatibility
4. economic availability, storage
5. the limiting nutrient concept
6. metal cofactors
lot to lot variation and quality control |
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Term
| why do cell culture media for growth of mammalian or insect cells require buffering in the physiological range of pH? what buffers are commonly used? |
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Definition
cell line formation causes formation of CO2 (pH rises) and lactic acid causes pH drop
Hepes buffer and exogenous CO2 are used |
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Term
| how do you determine the QC criteria for medium containing complex materials such as bovine serum? |
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Definition
low level of antibodies and growth factors, looking for filtration (0.l1 micrometre membrane filters) certificate of analysis, origin
cell growth performance testing, microbial sterility testing, screening for viruses endotoxins etc |
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Term
| What are 6 types of microbial or cell growth media used in biomanufacturing? |
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Definition
1. chemically defined
2. semi-defined
3. complex
4. reducing
5. achorage dependent cell culture
6. suspension cell culture |
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Term
| stoichiometry of microbial growth is determined by ___, ____, ____ |
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Definition
| cell composition, carbon source, products |
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Term
| what are two examples of C1 substrates |
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Definition
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Term
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Definition
| the substrate concentration at umax/2 |
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Term
| briefly explain how substrate concentration (S) determines the maximum cell density or in other words the limiting nutrient concept |
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Definition
| grow a shake flask on defined media and plot growth optical density versus concentration of limiting nutrient determine linear region of respons |
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