单链PEG化重组人干扰素ω的结构和性能表征  

Structure and Function of Mono-PEGylated Recombinant Human Interferon Omega

作　　者：潘红春[1,2] 杨琴[1,2] 苏秋菊[1,2] 刘丽[1,2] 刘红[1,2] 

机构地区：[1]西南大学药学院,重庆400716 [2]重庆药物过程与质量控制工程技术研究中心,重庆400716

出　　处：《西南大学学报（自然科学版）》2014年第2期7-13,共7页Journal of Southwest University(Natural Science Edition)

基　　金：国家重大新药创制项目(2010ZX09401-306-1-4);重庆市制药过程与质量控制工程技术研究中心能力提升项目(CSTC2012gg-yyjsb10002-33);西南大学博士基金项目(SWU110056和SWU110057)资助

摘　　要：用紫外扫描光谱、MALDI-TOFMS等电聚焦电泳、毛细管电泳、圆二色光谱、肽图等手段对单链PEGrhIFNω的性能、组分和结构进行表征．结果表明，单链PEG-rhIFNω的最大紫外吸收波长未改变，为280．6nm；等电点由9．7下降到7．35～8．60；质谱分析发现，单链PEG-rhIFNω中仍含有PEG多点修饰组分；圆二色光谱分析发现，单链PEG-rhIFNω二级结构中同时具有典型α螺旋和无规卷曲结构，而rhIFNm没有无规卷曲结构；毛细管电泳分析发现，单链PEG-rhIFNω含4种位置异构体．肽图分析推测，单链PEG-rhIFNω的PEG修饰位点应为Lys33或Lys167-和Lys46-或Lys52-或Lys152-残基．To characterize the the structures, components and properties of mono-PEOylated recombinant human interferon omega （mono-PEG-rhIFNω）, comparative studies of mono-PEG-rhIFNco and rhIFNω were performed by UV scanning spectrum, matrix-assisted laser desorption ionization time-of-flight mass spectrometry （MALDI-TOF MS）, isoelectric focusing electrophoresis （IEF）, capillary electrophoresis （CE）, circular dichroism （CD）, and peptide mapping analysis. The results showed that the maximum of UV absorption wavelength of mono-PEG-rhlFNω remained unchanged, being 280.6 nm, in good agree ment with 280.5 nm of rhlFNω, and the isoelectrie point was reduced from 9.7 for rhIFNω to 7.35--8.60 for mono PEG-rhIFNω. In MALDI TOF MS analysis, multi-PEGylated rhIFNω were detected in the mono PEG-rhIFNω. In CD analysis,α-helix and random coil were simultaneously observed in the seconda ry structure of mono-PEG-rhIFNω, while no random coil was detected in rhIFNω. Four positional isomers of mono-PEG-rhIFNco were discovered by CE analysis. It is speculated based on the peptide mapping anal- ysis of the mono-PEG-rhlFNω and rhlFNω samples treated by cyanogen bromide that PEGylation sites of the four positional isomers of mono-PEG-rhIFNω should be Lys33- or Lys167- and Lys46- or Lys52 or Lys152-residues .

关 键 词：重组人干扰素ω 聚乙二醇化 二级结构 位置异构体 肽图 

分 类 号：O632.14[理学—高分子化学]