超高效纳升液相色谱-电喷雾串联质谱鉴定重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白  

Characterization of Recombinant Human Necrosis Factor-α Receptor Ⅱ∶IgG Fc Fusion Protein by Nano Ultra-high Performance Liquid Chromatography-Electrospray Ionization Mass Spectrometry Tandem Mass Spectrometry

作　　者：柏兆方[1] 林碧蓉 李萍[1] 李卫华[1] 刘炳玉[1] 王红霞[1] 

机构地区：[1]国家生物医学分析中心,北京100850 [2]上海中信国健药业有限公司,上海201301

出　　处：《分析化学》2009年第7期1025-1028,共4页Chinese Journal of Analytical Chemistry

基　　金："十一五"国家科技支撑计划(No.2006BAF07B01)资助

摘　　要：采用超高效纳升液相色谱-电喷雾串联质谱对重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白进行了分析。样品经胰蛋白酶酶切后,进行液相色谱-串联质谱分析和数据库检索。结果表明,分别有5个和7个肽段匹配于人肿瘤坏死因子受体和人Iggamma-1chainCregion。比对分析表明,重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白中检测到的人IgG的7个肽段完全与人IgG1的序列匹配,与其它3个亚型只有部分肽段匹配。说明重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白一级结构正确,Fc片段确实为人IgG1的Fc片段。The Nano ultra-high performance liquid chromatography-electrospray ionization mass spectrometry tandem mass spectrometry（UPLC-ESI-MS/MS） was used to characterize the primary structure of recombinant human necrosis factor-α receptor Ⅱ： IgG Fc fusion protein. Samples were digested by trypsin, followed by liquid chromatography-tandem mass spectrometry analysis and database search. Five peptides matched with tumor necrosis factor receptor and seven peptides matched with human IgGl fragment of the Fc. Further analysis demonstrated that seven peptides all matched with the IgGl Fc but only partly peptides matched with the other subtypes. RhTNFR：Fc is fused by IgGl Fc but not other subtypes.

关 键 词：基因重组受体-抗体融合蛋白 纳升液相色谱 电喷雾串联质谱 蛋白鉴定 

分 类 号：R593.22[医药卫生—内科学]