Recombinant expression of proteins in a disulfide-bridged, two-chain form

Abstract

The invention relates to a method for producing polypeptides or proteins in a two-chain form by recombinant expression in E. coli host cells, whereby i) the polypeptide or protein exerts its biological activity as a two-chain polypeptide or protein, ii) the C-terminal amino acid group of the first chain is an Arg or Lys group, iii) the second chain of the protein / polypeptide, at its N terminus, has 1 to 20 amino acid groups and a pentapeptide sequence VPXGS referred to as PRS, wherein X may be any naturally occurring amino acid, wherein V represents Val, Leu, Ile, Ala, Phe, Pro or Gly, P represents Pro, Leu, Ile, Ala, Phe, Val or Gly, G represents Gly, Leu, Ile, Ala, Pro, Phe or Val and S represents Ser, Tyr, Trp or Thr.

Description

[0001] The application of the present invention is based on the application date of January 20, 2006, the application number "200680002957.3" (the international application number is PCT / DE2006 / 000088), and the name is "recombinant expression of protein in disulfide bridged double-chain form" A divisional application of an invention patent application. technical field [0002] One aspect of the invention relates to a method of producing a protein in double-chain form by recombinant expression in an E. coli host cell. Another aspect of the present invention relates to the double-chain biologically active form of the protein or polypeptide, which can be produced by the aforementioned method. [0003] Compared with corresponding recombinant proteins / polypeptides which do not exhibit the features of the present invention, they have the important advantage that no specific protease treatment is necessary for directed cleavage of the polypeptide chain, thus significantly simplifying...

AI Technical Summary

Problems solved by technology

The use of this endoprotease has two main disadvantages: On the one hand, it cannot be ruled out in any case that, in addition to the cleavage sites added by genetic technology methods, there are other added cleavage sites in the amino acid sequence site

Even when the cleavage efficiency of these minor cleavage sites is significantly lower, the mixture of different cleavage variants of the toxin can lead to separation difficulties after protease treatment

On the other hand, in the case of pharmaceutical preparations, for reasons of pharmaceutical law (regulatory considerations), the subsequent addition of proteins or exposure of the preparation to other proteins is very disadvantageous, since the protein and its possible contamination must be demonstrated substances are completely removed in further processing; this usually requires considerable expense

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