Production and recovery of recombinant neurotrophins

C - Chemistry – Metallurgy – 12 – N

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C12N 15/62 (2006.01) A61K 38/18 (2006.01) C07K 14/245 (2006.01) C07K 14/475 (2006.01) C07K 14/48 (2006.01) C07K 19/00 (2006.01) C12N 15/18 (2006.01) C12N 15/34 (2006.01) C12N 15/70 (2006.01)

Patent

CA 2111110

2111110 9222665 PCTABS00018 We disclose a process for producing and recovering a recombinant neurotrophin comprising the steps of: (1) culturing a non-animal host cell transformed with a recombinant DNA molecule comprising an expression control sequence operatively linked to a DNA sequence encoding a neurotrophin; (2) solubilizing the neurotrophin in a solution comprising a protease inhibitor in an amount effective to inhibit degradation of the neurotrophin by proteases and a strong denaturing agent, said solution being essentially free of reducing agents; (3) exchanging the strong denaturing agent for a weak denaturing agent; (4) adjusting the solution to comprise a basic amino acid at a concentration effective to maintain the solubility of the neurotrophin in a nondenaturing environment; (5) removing the weak denaturing agent; and (6) purifying the neurotrophin. In a specific embodiment, also disclosed is the bacterial production of recombinant neurotrophin-4.

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