title: Synthesis and purification of mono-PEGylated insulin.
abstract: 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.
authors: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China.
authors: Huaizhi
authors: H
authors: Dou
authors: Min
authors: M
authors: Zhang
authors: Yu
authors: Y
authors: Zhang
authors: Chunhua
authors: C
authors: Yin
mesh_terms: D000818:Animals; D007328:Insulin / Q000737:chemistry* / Q000302:isolation & purification* / Q000494:pharmacology; D051379:Mice; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D007328:Insulin; D011092:Polyethylene Glycols; C028210:monomethoxypolyethylene glycol