金黄色葡萄球菌TraP蛋白单克隆抗体制备  

Preparation of Monoclonal antibodies against Staphylococcus aureus TraP

作　　者：周雪[1] 范志勇 汤炜[1] 张丽萌[1] 李闰婷[1] 周瑜[1] 崔玉东[1] 

机构地区：[1]黑龙江八一农垦大学生命科学技术学院,大庆163319 [2]大庆市第四中学

出　　处：《黑龙江八一农垦大学学报》2012年第2期48-51,99,共5页journal of heilongjiang bayi agricultural university

基　　金：国家自然科学基金(项目编号31072120)

摘　　要：研究通过骨髓瘤细胞SP2/0与经金黄色葡萄球菌(S.aureus)TraP蛋白免疫的小鼠脾细胞融合,并进一步筛选和单克隆化,获得了11株稳定分泌抗TraP蛋白的单克隆抗体(McAb)杂交瘤细胞株,其中8株McAb(2A1、3A6、3B1、1B4、2C5、4C7、5C3和2D8)亚类为IgG1,3株McAb(2A7、3A1和1C4)亚类为IgM,轻链均为κ链。8株IgG1型McAb的小鼠腹水的抗体效价均达到1∶128 000,交叉实验结果显示这8株McAb不与S.aureus的ClfA、ClfB、Cna、FnbPA和IsdB蛋白以及链球菌的GapC蛋白反应,具有良好的特异性,Western-blotting结果显示这8株McAb识别的都是TraP的线性表位。Eleven antibody-secreting hybridomas were produced in vitro by fusing myeloma cells line SP2/0 with spleen cells from a BALB/c mice immunized with Staphylococcus aureus Target of RNAIII-activiting protein （TraP）. Eight （2A1,3A6,3B1,1B4,2C5, 4C7,5C3 and 2D8）of these monoelonal anti-TraP antibodies were characterized as subtype IgG1 and K chain, and the antibody titers of McAb produced by these eight hybridomas in ascites were 1：128 000. These McAbs specific for TraP were identified by an indirect ELISA against protein ClfA,ClfB,Cna,FnbPA and IsdB from Staphylococcus aureus and GapC from Streptococcus. Results of western-blotting suggested that these McAbs were the lined epitopes of TraP.

关 键 词：金黄色葡萄球菌 TRAP 单克隆抗体 

分 类 号：Q71[生物学—分子生物学]