金葡菌肠毒素C2的单克隆抗体制备、鉴定及初步应用  

Preparation and preliminary application of monoclonal antibody against Staphylococcal enterotoxin C2

作　　者：骆鹏[1] 吕慧贤[1] 张庶民[1] 

机构地区：[1]中国药品生物制品检定所,北京100050

出　　处：《中国免疫学杂志》2007年第1期53-55,共3页Chinese Journal of Immunology

摘　　要：目的：制备抗金葡菌肠毒素C2（SEC2）的单克隆抗体（MeAb），并建立SEC2检测方法。方法：以金葡菌培养液中纯化的SEC2为抗原，免疫BALB／c小鼠，制备单克隆抗体；并对单克隆抗体的特性进行鉴定；利用纯化后的抗体建立了夹心ELISA定量检测SEC2方法，并进行了验证和初步应用。结果：筛选出4株稳定分泌抗SEC2抗体的杂交瘤细胞株，其免疫球蛋白亚类均为IgG1，除两株单抗的SEC2识别位点相同外，其余单抗均特异性识别SEC2的不同结合位点。建立的夹心ELISA定量检测法，特异性、灵敏度、重复性均较好，检测SEC2范围为0．5—20ng,／ml，回收率在97．8％-101％，变异系数为2％-5％。结论：本研究制备的抗SEC2的单克隆抗体，可用于建立了SEC2定量检测方法，为控制金葡素制品质量和金葡菌肠毒素研究提供了较实用的方法。Objective：To obtain moneelonal antibodies（McAb） against Staphylococcal enterotoxin C2（SEC2） and establish method for detecting SEC2. Methods：The secreted SEC2 from staphylococcus aureus was used as antigen to immune BALB/c mice. Monoclonal antibodies against SEC2 were prepared by normal hybridoma technique. By identifying the characters of McAbs, the quantitative detection ELISA test method were established and were preliminarily applied. Results：Four hybridmas producing antibodies against SEC2 were obtained. IgG isotypes of four McAha were IgG1. Their binding site was different except two McAbs that shared the same binding site. The McAbs were proved to be specific for SEC2. The sandwich ELISA method had good specificity, sensitivity and reproducibility, and it was founded to be able to detect SEC2 at concentration from 0. 5 to 20 ng/ml. Its recovery ranged between 97. 8% and 101% ,and CV value ranged between 2% and 5%. Conclusion：The prepared McAb against SEC2 can be used for SEC2 immunoassay. This work provides a basis for controlling the quality of JINPUSU and researching staphylococcal enterotoxin.

关 键 词：金葡菌肠毒素C2 单克隆抗体 夹心ELISA 

分 类 号：R392.11[医药卫生—免疫学]