液质联用技术鉴定rLTB的结构  

The Structure Identification of rLTB by HPLC-MS

作　　者：杨珺[1] 田文标[1] 朱永红[1] 邹全明[1] 

机构地区：[1]中国人民解放军第三军医大学临床微生物教研室,重庆400038

出　　处：《解放军药学学报》2004年第3期174-177,共4页Pharmaceutical Journal of Chinese People's Liberation Army

基　　金："8 63"生物与现代农业技术领域生物工程技术主题课题;No .2 0 0 1AA2 15 161

摘　　要：目的　利用电喷雾离子化为接口 (ESI)的高效液相色谱 /质谱联用技术 (HPLC -MS) ,鉴定重组大肠杆菌不耐热肠毒素B亚单位 (rLTB)氨基酸序列结构的正确性。方法　利用SDS -PAGE分离、呈现蛋白混合物 ,胰蛋白酶原位水解rLTB ,HPLC -MS联机制备肽指纹图 ,分析肽指纹图获得rLTB氨基酸全序列信息。结果　rLTB的全序列结构与理论的全序列结构完全一致。结论　改进的HPLC -MS测定条件稳定 ,提高了质谱灵敏度 ,结合肽指纹图的分析结果 。Aim The correctness of the primary structure of recombinant heat-labile enterotoxin B was identified by liquid chromatography/electrospray ionization mass spectrometry.Methods rLTB in SDS-PAGE gel was digested by trypsin,and its peptide finger mapping was prepared by improved HPLC-MS methods.By analyzing peptide finger mapping,the primary structure of rLTB was identified.Results Theoretic and actual primary structure of rLTB is consistent.Conclusion Improved HPLC-MS conditions were stable.The sensitivify of MS was increased.Whole amino acid sequence of proteins were detected.

关 键 词：液质联用技术 鉴定 rLTB 大肠杆菌不耐热肠毒素B亚单位 DNA重组技术 指纹图谱 

分 类 号：R378.21[医药卫生—病原生物学]