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用单克隆抗体通过免疫沉淀和蛋白质印迹转移检测淋病奈瑟菌外膜蛋白III的抗原性。

Antigenicity of Neisseria gonorrhoeae outer membrane protein(s) III detected by immunoprecipitation and Western blot transfer with a monoclonal antibody.

作者信息

Swanson J, Mayer L W, Tam M R

出版信息

Infect Immun. 1982 Nov;38(2):668-72. doi: 10.1128/iai.38.2.668-672.1982.

Abstract

Some properties of gonococcal outer membrane III were studied by using a monoclonal antibody (2E6) in Western blot transfer and immunoprecipitation reactions. By Western blot, the 2E6 monoclonal had a single antigenic target: outer membrane protein III. All three gonococcal strains examined exhibited identical reactivities for their protein(s) III with this monoclonal. When 125I-labeled gonococci were incubated with monoclonal antibody 2E6, lysed in Zwittergent, and antibody-antigen complexes were immunoprecipitated with protein A-Sepharose, both outer membrane proteins I and III were present in the immunoprecipitates regardless of the gonococcal strain used. Exposure of gonococci to Iodogen and 2-mercaptoethanol influences the electrophoretic migration characteristics of protein(s) III in polyacrylamide gel electrophoresis with sodium dodecyl sulfate; the antigenicity of protein(s) III was unaltered when assessed by Western blot transfer after reaction with Iodogen or 2-mercaptoethanol. These data demonstrate that outer membrane protein III is a surface-exposed antigenic moiety common to diverse strains of gonococci.

摘要

利用单克隆抗体(2E6)通过蛋白质印迹转移和免疫沉淀反应研究了淋球菌外膜III的一些特性。通过蛋白质印迹法,2E6单克隆抗体有一个单一的抗原靶点:外膜蛋白III。所检测的三株淋球菌的蛋白III与该单克隆抗体表现出相同的反应性。当用125I标记的淋球菌与单克隆抗体2E6孵育,在两性离子去污剂中裂解,并用蛋白A-琼脂糖免疫沉淀抗体-抗原复合物时,无论使用何种淋球菌菌株,免疫沉淀物中均存在外膜蛋白I和III。将淋球菌暴露于碘昔芬和2-巯基乙醇会影响十二烷基硫酸钠聚丙烯酰胺凝胶电泳中蛋白III的电泳迁移特性;在用碘昔芬或2-巯基乙醇反应后通过蛋白质印迹转移评估时,蛋白III的抗原性未改变。这些数据表明外膜蛋白III是不同淋球菌菌株共有的表面暴露抗原部分。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a799/347790/1e3b06f5a7da/iai00146-0281-a.jpg

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