Process and compositions for the isolation of human relaxin

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

C - Chemistry, Metallurgy

12

N

C12N 15/16 (2006.01) C07K 1/12 (2006.01) C07K 14/64 (2006.01) C12N 1/21 (2006.01) C12P 21/00 (2006.01)

Patent

CA 2051375

A process is provided for cleaving a polypeptide into at lease 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.

L'invention concerne un procédé de clivage d'un polypeptide en au moins deux composantes du polypeptide, consistant à traiter une forme réduite, exempte de cystéine, du polypeptide, à l'aide d'un agent de clivage, dans des conditions permettant de cliver ledit polypeptide au niveau d'une jonction voulue entre les produits de clivage dudit polypeptide. Le procédé de clivage consiste de préférence à cultiver des cellules contenant de l'ADN codant ledit polypeptide, au moins un codon Asp se trouvant dans ledit ADN au niveau d'une jonction voulue entre les composantes à cliver les une des autres, ladite culture permettant d'obtenir l'expression de l'ADN pour produire ledit polypeptide dans la culture cellulaire hôte; on traite ensuite une forme réduite exempte de cystéine dudit polypeptide à l'aide d'acide dilué dans des conditions permettant de cliver ledit polypeptide au niveau de la jonction Asp. Dans des modes de réalisation particuliers, on a prévu une séquence d'ADN codant un précurseur de relaxine et comprenant des codons codant des liaisons contenant de l'acide aspartique au niveau de nouvelles positions dans le précurseur, ce qui facilite le clivage de peptides de relaxine A par traitement à l'aide d'acide dilué.

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