Enhanced purification and expression of insoluble recombinant pr

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

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43525233, 43525421, 530418, 530420, C12N 115, C12N 121, C12N 1509, C07K 100

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055232159

ABSTRACT:
A method for isolating a desired protein which comprises: providing a host cell expressing the desired protein as an insoluble aggregate; culturing the host cell with an effective mount of Cu.sup.++ ; disrupting the host cell producing a lysate; incubating the insoluble fraction non-disulfide-bond reducing or non-copper competing chaotropic conditions to solubilize contaminants; separating the insoluble from the soluble fraction; and exposing the insoluble fraction to disulfide-bond reducing or copper competing chaotropic conditions to solubilize the desired protein.

REFERENCES:
patent: 4656131 (1987-04-01), Kitano
patent: 4748234 (1988-05-01), Dovin
patent: 5342921 (1994-08-01), Cousens
Mitraki et al., Protein Folding Intermediates and Inclusion Body Formation (1989) Biotechnology 7: 690-697.
Georgiou, G., AIChE Journal, 34(8):1233-1248 (1988).

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