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目的:建立检测聚乙二醇位点特异性修饰重组人干扰素α-2b反应的方法。方法:采用分子量20000的甲氧基聚乙二醇马来酰亚胺修饰重组人干扰素α-2b,反应混合物样品经十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)后,碘染色法判断反应产物组成。结果:该修饰反应产物除含有单PEG化的干扰素α-2b外,还有不同修饰程度的多PEG化干扰素。结论:本方法方便快捷、分辨率高、特异性强,同时可用于其它聚乙二醇修饰蛋白质的分析研究。
OBJECTIVE: To establish a method for detecting the site-specific modification of recombinant human interferon α-2b by polyethylene glycol. METHODS: Recombinant human interferon α-2b was modified by methoxypolyethylene glycol maleimide with a molecular weight of 20000. After the sample of the reaction mixture was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) Iodine staining to determine the reaction product composition. Results: In addition to mono-pegylated interferon alpha-2b, the modified reaction product also has different degree of modification of poly-pegylated interferon. Conclusion: The method is convenient, rapid, high resolution and specificity, and can be used for other polyethylene glycol modified protein analysis.