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biotech chapter 3

biotechnology cluster 2

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