胰高血糖素样肽-1的聚乙二醇定点修饰
Investigation on Site-specific PEGylation of Glucagon-like Peptide-1
查看参考文献11篇
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文摘
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采用分子量为5 kDa的单甲氧基聚乙二醇琥珀酰亚胺碳酸酯(mPEG-SC)修饰胰高血糖素样肽-1(GLP-1),建立了GLP-1的定点修饰反应及分离纯化工艺,考察了修饰反应各因素对单修饰产物得率及体外活性的影响,得到优化修饰条件为:pH 7.5, 50 mmol/L Na_2HPO_4-NaH_2PO_4缓冲液,GLP-1浓度1 mg/mL, PEG/GLP-1摩尔比2:1,4℃下反应1 h.在此条件下,单修饰PEG-GLP-1得率达50%.反相高效液相色谱分离纯化所得单修饰产品纯度达98%,体外活性保留为未修饰GLP-1的86%,其在Sprague-Dawley远交群大鼠体内的循环半衰期为60 min,比原肽提高了20倍. |
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其他语种文摘
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In order to obtain a more stable PEGylated glucagon-like peptide-1, and prolong its half life, glucagon-like peptide-1 was site-specifically modified with monomethoxy polyethylene glycol succinimidyl carbonate (mPEG-SC, 5000 kDa). It was found that the optimized reaction conditions for maximized bioactivity and the highest modification yield of mono PEGylated GLP-1 were as follows: in 50 mmol/L and pH 7.5 phosphate buffer, concentration of GLP-1 1.0 mg/mL, molar ratio of PEG to GLP-1 2:1, and reaction time 1 h at 4℃. Under the optimized conditions, the mono modification yield reached 50%. Reverse phase high performance liquid chromatography was used to separate and purify mono PEGylated GLP-1 from the reaction mixture. The purity of mono PEGylated glucagon-like peptide-1 was higher than 98%, characterized by gelfiltration chromatography, the bioactivity of mono PEGylated glucagon-like peptide-1 was 86% of the native GLP-1, while its half life in Sprague-Dawley rats was much longer than that of native GLP-1. The mono PEGylated glucagon-like peptide-1 was also stable in aqueous solution. |
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来源
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过程工程学报
,2009,9(6):1169-1173 【核心库】
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关键词
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胰高血糖素样肽-1
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聚乙二醇
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定点修饰
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地址
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中国科学院过程工程研究所, 生化工程国家重点实验室, 北京, 100190
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语种
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中文 |
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文献类型
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研究性论文 |
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ISSN
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1009-606X |
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学科
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生物化学;药学 |
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基金
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国家自然科学基金
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国家863计划
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北京市自然科学基金
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文献收藏号
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CSCD:3763597
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