PRODUCTION AND RECOVERY OF RECOMBINANT NEUROTROPHINS

摘要

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.