Processes and compositions for the isolation of human relaxin

Details Processes and compositions for the isolation of human relaxin Processes and compositions for the isolation of human relaxin

1992-07-01

1995-11-07

Chambers, Jasemine C.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

435 692, 435 697, 435 701, 435 703, 43525233, 435849, 530344, 530345, 530350, 530427, 530300, 930240, 930310, 930DIG534, 935 73, C12P 2106, C12P 2102, C12N 1570, C12N 1512

Patent

active

054647568

ABSTRACT:
A process is provided for cleaving a polypeptide into at least two polypeptide components comprising treating a reduced, free-cysteine form of the polypeptide with a cleaving agent under conditions for cleaving the polypeptide at a desired junction between the polypeptide cleavage products. More preferably, the process for cleaving comprises culturing cells containing DNA encoding said polypeptide, wherein at least one Asp codon is present in said DNA at a desired junction between the components to be cleaved from each other, said culturing resulting in expression of the DNA to produce the polypeptide in the host cell culture; and treating a reduced, free-cysteine form of the polypeptide with dilute acid under conditions for cleaving the polypeptide at the Asp junction. In particular embodiments, a DNA sequence is provided that encodes a relaxin precursor and includes codons encoding aspartic acid-containing linkers at novel positions within the precursor, allowing the ready cleavage of relaxin A peptides by treatment with dilute acid.

REFERENCES:
patent: 4758516 (1988-07-01), Hudson et al.
patent: 4835251 (1989-05-01), Burnier et al.
patent: 4871670 (1989-10-01), Hudson et al.
Inglis, 1983, Methods in Enzymology 91:324-332.
Landon et al. Meth. Enzymol. 1977 (47):145.
Nishikawa et al. 1987 Protein Eng 1(6):487, Abstract.

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