Method for recovery of recombinantly produced polypeptides involving the cleavage of fusion proteins

摘要

A method for the preparation of a recombinant polypeptide comprising a) Introducing into a host cell an expression vector comprising: 1) A nucleic acid sequence capable of regulating transcription into a host cell, operatively linked to 2) A chimeric nucleic acid sequence encoding a fusion protein, the chimeric nucleic acid sequence comprising (a) A nucleic acid sequence encoding a pro-peptide derived from an autocatalytically maturing zymogen, linked in reading frame to (b) a nucleic acid sequence heterologous to the pro-peptide and encoding the recombinant polypeptide, wherein the heterologous nucleic acid sequence is located immediately downstream of the nucleic acid sequence encoding the pro-peptide; operatively linked to 3) A nucleic acid sequence encoding a termination region functional in said host cell b) Growing the host cell to produce said fusion protein; and c) Altering the environment of the fusion protein so that the pro-peptide is cleaved from the fusion protein to release the recombinant polypeptide, but excluding a human host cell in situ. Altering the environment can take place under in vitro or in vivo conditions. The fusion protein produced can be used to prepare pharmaceutical compositions or food compositions. The medicaments prepared are for delivering a therapeutic or nutritional protein to a human or animal.