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›› 2010, Vol. 10 ›› Issue (3): 593-597.

• 生化工程专栏 • Previous Articles     Next Articles

Modification of Glucagon-like Peptide-1 in Ethanol with Polyethylene Glycol

WANG You-ao, ZHAI Yan-qin LEI Jian-du MA Guang-hui SU Zhi-guo   

  1. State Key Lab of BiochemicaL Engineering, Institute of Process Engineering, Chinese Academy of Sciences Multi-phase Reaction Laboratory, Institute of Process Engineering Chinese Academic of Science National Laboratoru of Biochemical Engineering, Insitute of Process Engineering National Laboratoru of Biochemical Engineering, Insitute of Process Engineering National Laboratoru of Biochemical State Key Lab. Biochem. Eng., Inst. Process Eng., Chinese Academy of Sciences State Key Lab. Biochem. Eng., Inst. Process Eng., Chinese Academy of Sciences
  • Received:2010-03-30 Revised:2010-05-14 Online:2010-06-20 Published:2010-06-20
  • Contact: WANG You-ao,

胰高血糖素样肽-1在乙醇中的聚乙二醇修饰

王友傲 翟艳琴 雷建都 马光辉 苏志国   

  1. 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所生化工程国家重点实验室
  • 通讯作者: 王友傲

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Abstract: Glucagon-like peptide-1 (GLP-1) was modified with monomethoxy polye(ethylene glycol succinimidyl carbonate) by using ethanol as a reaction medium. The reaction conditions were investigated and optimized, and the separation procedures of product Mono-PEG-GLP-1 were developed. After that, the stability in vitro and activity in vivo of Mono-PEG-GLP-1 were investigated. The optimized reaction conditions were obtained as follows: concentration of GLP-1 1.0 mg/mL, molar ratio of PEG to GLP-1 1:1, and reaction time 24 h at 37℃. Under the optimized conditions, the yield of Mono-PEG-GLP-1 was up to 77%. The Mono-PEG-GLP-1 was separated from reaction mixture by a preliminary centrifugation under frozen temperature and successive purification by reverse phase chromatography. The purity of Mono-PEG-GLP-1 was higher than 97%, as characterized by size exclusion chromatography. The Mono-PEG-GLP-1 showed better stability in plasma, decreased proteolysis to trypsin and better activity in vivo.

Key words: glucagon-like peptide-1, monomethoxy poly(ethylene glycol succinimidyl carbonate), PEGylation, ethanol

摘要： 以乙醇为溶剂，单甲氧基聚乙二醇琥珀酰亚胺碳酸酯(mPEG-SC)为修饰剂，对胰高血糖素样肽-1(GLP-1)进行了修饰反应条件的优化，同时对单修饰产物Mono-PEG-GLP-1进行分离纯化，并考察了其体外稳定性和体内活性. 得到的优化反应条件为：GLP-1浓度1 mg/mL, mPEG-SC与GLP-1摩尔比1:1, 37℃下反应24 h，该产物最高转化率达77%. 采用冷冻离心方式对Mono-PEG-GLP-1进行初步分离和浓缩，然后经反相液相色谱进一步高效纯化，纯度可达97%以上. 体外实验表明，Mono-PEG-GLP-1在血清中具有更好的稳定性及更强的抗胰蛋白酶消化能力. 体内动物实验表明，Mono-PEG-GLP-1具有更好的降血糖作用.

关键词: 胰高血糖素样肽-1, 单甲氧基聚乙二醇琥珀酰亚胺碳酸酯, 聚乙二醇修饰, 乙醇

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