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作者介绍了表现有沙眼衣原体血清型特异性(血清型A、B-Ba 或C)和血清组特异性(B、中间或C 血清组)的单克隆抗体的制造和定性。衣原体在HeLa229细胞中生长,原生小体(EB)用Renografin 密度梯度离心纯化。用活EB 为抗原作斑点免疫印迹测定筛选杂交瘤上清。用Western 印迹筛选证实其与主要外膜蛋白(MOMP)结合时为阳性上清。选取在两种测定中表现免疫反应性的单克隆抗体(McAb)。然后测定McAb 中和衣原体对小鼠毒性的能力。将5μ1McAb(100μg蛋白)与悬浮在1ml 蔗糖-磷酸盐-谷氨酸
The authors describe the manufacture and characterization of monoclonal antibodies that exhibit serotype specific (serotype A, B-Ba or C) and serogroup specific (B, intermediate or C serogroups) of C. trachomatis. Chlamydia were grown in HeLa229 cells, and primary bodies (EBs) were purified by Renografin density gradient centrifugation. Hybridoma supernatants were screened using a live EB antigen as a dot immunoblot. Screening with Western blot confirmed positive supernatant when bound to the major outer membrane protein (MOMP). Monoclonal antibodies (McAbs) that showed immunoreactivity in both assays were selected. The ability of McAb to neutralize Chlamydia in mice was then tested. Mu] M McAb (100 [mu] g protein) was suspended in 1 ml sucrose-phosphate-glutamic acid