Synthesis and Purification of Mono-PEGylated Insulin

Overview

Journal Chem Biol Drug Des

Specialties Biochemistry
Pharmacology

Date 2007 Mar 27

PMID 17381727

Citations 4

Authors

Huaizhi Dou

Min Zhang

Yu Zhang

Chunhua Yin

Affiliations

Soon will be listed here.

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.

Citing Articles

Trehalose Glycopolymer Enhances Both Solution Stability and Pharmacokinetics of a Therapeutic Protein.

Liu Y, Lee J, Mansfield K, Ko J, Sallam S, Wesdemiotis C Bioconjug Chem. 2017; 28(3):836-845.

PMID: 28044441 PMC: 5352466. DOI: 10.1021/acs.bioconjchem.6b00659.

Computational and nonglycosylated systems: a simpler approach for development of nanosized PEGylated proteins.

Mirzaei H, Kazemi B, Bandehpour M, Shoari A, Asgary V, Shafiee Ardestani M Drug Des Devel Ther. 2016; 10:1193-200.

PMID: 27042012 PMC: 4801162. DOI: 10.2147/DDDT.S98323.

Design, modeling, expression, and chemoselective PEGylation of a new nanosize cysteine analog of erythropoietin.

Cohan R, Madadkar-Sobhani A, Khanahmad H, Roohvand F, Aghasadeghi M, Hedayati M Int J Nanomedicine. 2011; 6:1217-27.

PMID: 21753873 PMC: 3131188. DOI: 10.2147/IJN.S19081.

Toward top-down determination of PEGylation site using MALDI in-source decay MS analysis.

Yoo C, Suckau D, Sauerland V, Ronk M, Ma M J Am Soc Mass Spectrom. 2008; 20(2):326-33.

PMID: 19019698 DOI: 10.1016/j.jasms.2008.10.013.