11.22 Fractionation of cell homogenate Recombinant insulin is produced using E.

Question

11.22 Fractionation of cell homogenate Recombinant insulin is produced using E. coli fermentation. The harvested cells are homogenised using a bead mill to release the protein inclusion bodies. The cell homogenate is then treated with CNBr to cleave the cell proteins into small fragments, leaving the proinsulin as a large, uncleaved molecule. The CNBr-treated homogenate contains several components, including 5% w/w cell debris, 8% w/w proinsulin, 5% w/w cleaved proteins, and 0.5% w/w sugar. The mixture is treated using membrane filtration to remove the smaller molecules and fragments. The membrane chosen for this application gives retention coefficients of 0.95 for the cell debris, 0.90 for proinsulin, 0.25 for cleaved protein, and 0 for sugar. The effect of fouling on solute retention is negligible. The cell debris is concentrated fivefold by this process. (a) What is the composition of the retentate? (b) What is the composition of the permeate?

Explanation / Answer

A)The retentate comprises mainly of cell debris in a concentrated form as it will be retained along with insulin and some part of cleaved proteins.

B)the permeate will have components that include mainly sugar long with the buffer employed for homogenization.

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