Method for making mature insulin polypeptides

摘要

The invention is related to a method for making human insulin analogs by culturing an fungi cell comprising a DNA vector encoding a precursor for human insulin analog, wherein the said precursor comprises a connecting peptide flanked with cleavage sites at both junctions with the A- and the B-chain of the insulin peptide, respectively said cleavage sites being cleaved within the fungi cell allowing the cell to secrete high amount of correctly processed, mature two chain human insulin analog to the culture media.

主权项

1. A method for making mature human insulin analogues by culturing a fungi cell comprising a DNA vector encoding a precursor for a human insulin analogue, wherein the said precursor comprises a connecting peptide flanked with cleavage sites at both junctions to the A- and B-chain of the insulin peptide, respectively, said cleavage sites being cleaved within the fungi cell allowing the cell to secrete the correctly processed mature, two chain human insulin analogue into the culture media, wherein the insulin analogue has Glu at position A14, and a mutation in at least one of the positions A8, B10, and A18 in the human insulin A- and/or the B-chain selected from the group consisting of Asp, Glu, His, Gln and Arg.