针对巨片形吸虫排泄分泌抗原的单克隆抗体的研制与特性分析

Development and characterization of monoclonal antibodies against excretory-secretory antigens of Fasciola gigantica.

作者信息

Viyanant V, Upatham E S, Sobhon P, Krailas D, Ardseungnoen P, Anatawara S

机构信息

Department of Biology, Faculty of Science, Mahidol University, Bangkok, Thailand.

出版信息

Southeast Asian J Trop Med Public Health. 1997;28 Suppl 1:128-33.

PMID:9656363

Abstract

Monoclonal antibodies (MAbs) directed against Fasciola gigantica excretory-secretory (ES) antigens were developed from BALB/c mice. Four were selected for further study, from the panel of hybridomas. The antigen specificities of these MAbs were characterized and localized by enzyme-linked immunoeletrotransfer blot (EITB) and immunoperoxidase technique. The target epitopes of these MAbs are 66 kDa protein (MAb 2D10), 66 and 27-26 kDa proteins (MAbs 5D10 and 4F5) and 27-26 kDa protein (MAb 2D9). MAb 2D9 reacted to the antigenic components of the luminal content and epithelial cell lining the cecum, whereas MAb 2D10 reacted specifically to the antigens of the tegument and surface membrane. It was found that all MAbs cross-reacted to various degrees with the antigens extracted from Schistosoma mansoni, S. mekongi, S. spindale and Paramphistomum spp. However, when MAbs were diluted to 1:100 or 1:400 significant reduction of their cross-reactivities was observed.

摘要

针对巨片形吸虫排泄分泌（ES）抗原的单克隆抗体（MAb）是从BALB/c小鼠中制备的。从杂交瘤细胞系中挑选出4种进行进一步研究。通过酶联免疫电转移印迹（EITB）和免疫过氧化物酶技术对这些单克隆抗体的抗原特异性进行了鉴定和定位。这些单克隆抗体的靶表位分别是66 kDa蛋白（单克隆抗体2D10）、66 kDa和27 - 26 kDa蛋白（单克隆抗体5D10和4F5）以及27 - 26 kDa蛋白（单克隆抗体2D9）。单克隆抗体2D9与盲肠腔内内容物和上皮细胞衬里的抗原成分发生反应，而单克隆抗体2D10则特异性地与皮层和表面膜的抗原发生反应。研究发现，所有单克隆抗体与从曼氏血吸虫、湄公血吸虫、间插血吸虫和双腔吸虫属提取的抗原均有不同程度的交叉反应。然而，当单克隆抗体稀释至1:100或1:400时，其交叉反应性显著降低。