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Site-Specific Chemical Modification of a Glycoprotein Fragment Expressed in Yeast

2025-03-27 生物化学 加入收藏
Site-specific modification of glycoproteins has wide application in both biochem

Site-specific modification of glycoproteins has wide application in both biochemical and biophysical studies. This method describes the conjugation of synthetic molecules to the N-terminus of a glycoprotein fragment, viz., human immunoglobulin G subclass 1 fragment crystallizable (IgG1 Fc), by native chemical ligation. The glycosylated IgG1 Fc is expressed in a glycosylation-deficient yeast strain. The N-terminal cysteine is generated by the endogenous yeast protease Kex2 in the yeast secretory pathway. The N-terminal cysteine is then conjugated with a biotin thioester to produce a biotinylated, glycosylated IgG1 Fc using native chemical ligation.

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