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biotech chapter 3
biotechnology cluster 2
| Question | Answer |
|---|---|
| Blotting used in making a recombinant protein | Southern Blotting |
| Best host cell for small simple proteins | bacteria and viruses |
| complex, glycosalated proteins would need what kind of cell | mammalian cells |
| batch culture | a few generations grow environment conditions get used up |
| fed batch culture | growth limiting nutrient |
| continuous culture and production | maintained under constant conditions with a constant nutrient supply the microorganism can grow steadily for a long period of time |
| stirred tank bioreactor | most reliable stirred to generate mixing for mass transfer stirred with paddles |
| airlift bioreactor | stirred with air flow instead of paddles |
| microcarrier bioreactor | fixed bed biorectors support matrix that allows for cells to grow carrier material may be: gelatin,collagen or cellulose |
| membrane bioreactor | hollow fiber perfusion uses membrane and bioreactor to cultivate and purify |
| Mediums that are used for cell culture | fetal calf/synthetic serum, vitamins, amino acids, carbon source, water, electrolytes (Ca, NA, K, PO4), trace minerals (Fe, Mg, Cu, Zn, etc.), and hormones (growth factor) |
| contaminates | viruses, bacteria, cellular DNA, protein |
| precautions | wear latex gloves, use sterilized glassware instead of baked, use treated water, clean all utensils |
| cdeal with contaminants by: | centrifugation, filtration, selective precipation and chromatography |
| scale-up: | number of processes used to isolate a certain protein produce a product at a high quality at a competitive price |
| Step1 of scale up: | particulate removal- removal of cell debris |
| step 2 of scale up: | concentration-take everything that was not removed and put it back together |
| step 3 of scale up: | protein capture-isolate the protein that you need (locate it) |
| step 4 of scale up: | removal of contaminants (final purification)-remove any contaminants |
| step 5 of scale up: | sterilization-finalize the product, prepare drug for the patient |
| size-exclusion chromatography | separates proteins based on their size |
| reverse phase chromatography or hydrophobic interaction chromatography | separates the proteins based upon the hydrophobic properties of some proteins |
| ion exchange chromatography | separates proteins based on their charges |
| affinity chromatography | how well a protein binds to a ligand |
| transgene | foreign gene placed in the embryo and the gene develops as the animal matures |
| pharm animal (transgenic) | animal that the transgene is inserted into it expresses the gene |
| inclusion bodies | nuclear or cytoplasmic aggregates of sustainable substances, usually proteins |
| why can inclusion bodies be problematic? | A lot of you product is being tied up in the inclusion bodies so you do not get the yield that you want |
Created by:
ljamison
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