Monoclonal antibodies against isoelectrically focused Nocardia asteroides proteins characterized by the enzyme-linked immunoelectro-transfer blot method.

Diagnostic immunology Pub Date : 1986-01-01
F A el-Zaatari, E Reiss, M A Yakrus, S L Bragg, L Kaufman
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Abstract

Sera from rabbits immunized with culture filtrates and homogenates of Nocardia asteroides B1042 gave at least eight precipitin bands by immunoelectrophoresis. At least 20 proteins with isoelectric points (pls) in the pH 4 to 5.4 range were observed in isoelectric focusing patterns. The enzyme-linked immunoelectro-transfer blot (EITB) assay showed that several of the isofocused proteins reacted with rabbit antisera and with sera from nocardiosis and tuberculosis patients. Antibodies against three proteins with pls of 4, 4.43, and 4.68 (antigenic factors 1,6,8) were present in nocardiosis patients' sera. The proteins were excised from isofocused gels, and IgG monoclonal antibodies (MAbs) were produced by the hybridoma method. MAbs against factors 1 and 6 did not crossreact with cytoplasmic antigens of Mycobacterium chelonae, M. intracellulare serotypes 4B and 8A, M. fortuitum, M. gordonae, or M. kansasii in the EITB method. Factor 8 (MAb) crossreacted with antigens of M. intracellulare and M. fortuitum.

用酶联免疫电转移印迹法鉴定抗等电聚焦诺卡菌小行星蛋白的单克隆抗体。
用小行星诺卡菌B1042培养滤液和匀浆免疫兔血清,免疫电泳结果显示至少有8条沉淀带。在pH值4 ~ 5.4范围内,至少有20种具有等电点(pls)的蛋白质以等电聚焦模式被观察到。酶联免疫电转移印迹(EITB)试验表明,几种同焦蛋白与兔抗血清、诺卡病和结核病患者血清发生反应。诺卡病患者血清中存在针对三种蛋白的抗体,其pls分别为4、4.43和4.68(抗原因子1,6,8)。从异聚焦凝胶中切除蛋白,通过杂交瘤法产生IgG单克隆抗体(mab)。在EITB方法中,针对因子1和6的单克隆抗体与chelonae、M.胞内血清型4B和8A、M. fortuitum、M. gordonae和M. kansasii的细胞质抗原没有交叉反应。因子8 (MAb)与M.胞内和M. fortuitum抗原发生交叉反应。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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