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用放射免疫分析法筛选抗曼氏血吸虫活童虫的鼠单克隆抗体。

Screening of murine monoclonal antibodies against living schistosomula of Schistosoma mansoni by radioimmunoassay.

作者信息

Wiest P M, Wisnewski A V, Johnson J H, Ramirez B, Kresina T F, Olds G R

机构信息

Program in Geographic Medicine, Brown University, Providence, RI.

出版信息

Int J Parasitol. 1991 Jul;21(4):449-54. doi: 10.1016/0020-7519(91)90102-d.

Abstract

A radioimmunoassay was developed to screen supernatants of murine monoclonal antibodies against surface antigens of living schistosomula of Schistosoma mansoni. Of 196 clones screened, 10% bound schistosomula. Of these, 74% bound only schistosomula. The remaining molecules also reacted with soluble adult worm antigens and soluble egg antigens as determined by enzyme-linked immunosorbent assay. Immunoblot analysis demonstrated that monoclonal antibody 204-3E4 reacted with a 68 kDa protein, a glycoprotein that induces substantial resistance against S. mansoni infection. Recognition of an 18 kDa antigen by 204-3F1 antibody was stage-specific with the antigen being expressed in cercariae, 3- and 24-h-old parasites but not 4-day, lung stage or adult worms. Monoclonal antibody 204-4E3 reacted with purified S. mansoni paramyosin. These data indicate that radioimmunoassay using living schistosomula is a rapid alternative method to identify murine hybridomas that secrete antibodies which react with surface antigens of S. mansoni.

摘要

开发了一种放射免疫测定法,用于筛选针对曼氏血吸虫活童虫表面抗原的鼠单克隆抗体的上清液。在筛选的196个克隆中,10%与童虫结合。其中,74%仅与童虫结合。通过酶联免疫吸附测定法确定,其余分子也与可溶性成虫抗原和可溶性虫卵抗原发生反应。免疫印迹分析表明,单克隆抗体204-3E4与一种68 kDa的蛋白质发生反应,该蛋白质是一种糖蛋白,可诱导对曼氏血吸虫感染的显著抗性。204-3F1抗体对18 kDa抗原的识别具有阶段特异性,该抗原在尾蚴、3小时和24小时龄的寄生虫中表达,但在4日龄、肺期或成虫中不表达。单克隆抗体204-4E3与纯化的曼氏血吸虫副肌球蛋白发生反应。这些数据表明,使用活童虫的放射免疫测定法是一种快速的替代方法,可用于鉴定分泌与曼氏血吸虫表面抗原发生反应的抗体的鼠杂交瘤。

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