可区分爱泼斯坦-巴尔病毒核抗原2A（EBNA2A）和2B（EBNA2B）的大鼠单克隆抗体。

Rat monoclonal antibodies differentiating between the Epstein-Barr virus nuclear antigens 2A (EBNA2A) and 2B (EBNA2B).

作者信息

Kremmer E, Kranz B R, Hille A, Klein K, Eulitz M, Hoffmann-Fezer G, Feiden W, Herrmann K, Delecluse H J, Delsol G, Bornkamm G W, Mueller-Lantzsch N, Grässert F A

机构信息

Institut für Immunologie, Hämatologikum, GSF, München, Germany.

出版信息

Virology. 1995 Apr 1;208(1):336-42. doi: 10.1006/viro.1995.1157.

DOI:10.1006/viro.1995.1157

PMID:11831716

Abstract

Rat monoclonal antibodies were produced against the C-terminus of Epstein-Barr virus nuclear antigens 2A (EBNA2A) and 2B (EBNA2B) expressed as bacterial trpE fusion proteins. The initial screening was performed using a soluble bacterial extract containing the fusion proteins. Positive hybridomas were confirmed by immunofluorescence on SF158 (Spodoptera frugiperda) insect cells infected with recombinant baculovirus (Autographa californica nuclear polyhedrosis virus) and expressing the complete EBNA2A or EBNA2B genes. We selected a panel of antibodies which reacted either with both antigens or specifically with EBNA2A or with EBNA2B. The antibodies were extensively characterized using immunoprecipitation, Western blotting, epitope mapping on synthesized peptide segments of EBNA2A, immunocytology, and immunohistology on both cryostat sections and paraffin sections of AIDS-associated primary central nervous system lymphomas.

摘要

制备了针对作为细菌trpE融合蛋白表达的爱泼斯坦-巴尔病毒核抗原2A（EBNA2A）和2B（EBNA2B）C末端的大鼠单克隆抗体。最初的筛选使用含有融合蛋白的可溶性细菌提取物进行。通过对感染重组杆状病毒（苜蓿银纹夜蛾核型多角体病毒）并表达完整EBNA2A或EBNA2B基因的SF158（草地贪夜蛾）昆虫细胞进行免疫荧光，确认阳性杂交瘤。我们选择了一组抗体，它们要么与两种抗原都反应，要么特异性地与EBNA2A或EBNA2B反应。使用免疫沉淀、蛋白质印迹、EBNA2A合成肽段上的表位作图、免疫细胞学以及对艾滋病相关原发性中枢神经系统淋巴瘤的冷冻切片和石蜡切片进行免疫组织学，对这些抗体进行了广泛表征。