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    • 2. 发明申请
    • Method for making maturated insulin polypeptides
    • 制备成熟胰岛素多肽的方法
    • US20100184133A1
    • 2010-07-22
    • US12311431
    • 2007-09-26
    • Per NorgaardAsser Sloth Andersen
    • Per NorgaardAsser Sloth Andersen
    • C12P21/06
    • C07K14/62
    • The invention is related to a method for making mature human insulin or an analogue thereof by culturing a fungi cell comprising a DNA sequence encoding a precursor for human insulin or an analogue of human insulin which precursor comprises the B-chain of human insulin or an analogue thereof, the A-chain of human insulin or an analogue thereof and a C-peptide linking the B-chain and the A-chain together thereof, wherein the C-peptide comprises at least one Kex2 cleavage site and an amino acid sequence attached at one end to the C-terminal amino acid residue in the B-chain and at the other end to the Kex2 site which amino acid sequence will facilitate a more efficient Kex2 cleavage within the fungi cell. The C-terminal extension of the B-chain may furthermore be capable of subsequently being cleaved off from the C-terminal amino acid residue in the B-chain by means of a carboxypeptidase activity either within the fungi cell or subsequently in the culture medium.
    • 本发明涉及通过培养包含编码人胰岛素前体的DNA序列或人胰岛素类似物的真菌细胞来制备成熟人胰岛素或其类似物的方法,该前体包含人胰岛素的B链或类似物 其中,人胰岛素的A链或其类似物和将β-链和A链连接在一起的C-肽,其中所述C-肽包含至少一个Kex2切割位点和附着于其上的氨基酸序列 一端到B链中的C-末端氨基酸残基,另一端连接到Kex2位点,该氨基酸序列将促进真菌细胞内的更有效的Kex2切割。 B链的C-末端延伸还可以随后通过真菌细胞内或随后在培养基中的羧肽酶活性随后从B链中的C末端氨基酸残基断裂。