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Synthesis and Purification of Mono‐PEGylated Insulin
Author(s) -
Dou Huaizhi,
Zhang Min,
Zhang Yu,
Yin Chunhua
Publication year - 2007
Publication title -
chemical biology and drug design
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.59
H-Index - 77
eISSN - 1747-0285
pISSN - 1747-0277
DOI - 10.1111/j.1747-0285.2007.00479.x
Subject(s) - ethylene glycol , insulin , aldehyde , chemistry , peg ratio , chromatography , high performance liquid chromatography , pegylation , polyethylene glycol , biochemistry , organic chemistry , medicine , catalysis , finance , economics
Monomethoxypoly(ethylene glycol) 5000 (mPEG5K) and 2000 (mPEG2K) were activated to produce mPEG‐aldehyde with content over 80%. Effects of reaction time, pH and initial molar ratio of mPEG‐aldehyde to insulin on synthesis of PEGylated insulin were studied, taking mPEG5K‐aldehyde as a model. On the optimized condition, approximately 90% of product was mono‐PEGylated insulin. Mono‐PEGylated insulin was purified by ion‐exchange chromatography. The purity of each component was analysed by RP‐HPLC and the results showed an efficient purification. Matrix assistant laser desorption ionization time‐of‐flight mass spectra demonstrated that the modification site of mono‐PEGylated insulin was PheB1. The biological activity of all PEGylated insulin was also evaluated.