CAJ | 학술논문

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Ig gamma-1 chain C region.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.
채용초고효납승액상색보-전분무천련질보대중조인Ⅱ형종류배사인자수체-항체융합단백진행료분석.양품경이단백매매절후,진행액상색보-천련질보분석화수거고검색.결과표명, 분별유5개화7개태단필배우인종류배사인자수체화인Ig gamma-1 chain C region.비대분석표명,중조인Ⅱ형종류배사인자수체-항체융합단백중검측도적인IgG적7개태단완전여인IgG1적서렬필배,여기타3개아형지유부분태단필배.설명중조인Ⅱ형종류배사인자수체-항체융합단백일급결구정학,Fc편단학실위인IgG1적Fc편단.
The Nano ultra-high performance liquid chromatography-electrospray ionization mass spectrometry tandem mass spectrometry(UPLC-ESI-MS/MS) was used to characterize the primary 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 IgG1 fragment of the Fc. Further analysis demonstrated that seven peptides all matched with the IgG1 Fc but only partly peptides matched with the other subtypes. RhTNFR:Fc is fused by IgG1 Fc but not other subtypes.