仿刺参体腔细胞单克隆抗体的制备及特性分析  

Preparation and characterization of monoclonal antibodies against coelomocytes of sea cucumber Apostichopus japonicus

作　　者：李强[1] 迟刚[1] 李莹[1] 李华[1] 

机构地区：[1]大连海洋大学农业部海洋水产增养殖学重点实验室,辽宁大连116023

出　　处：《大连水产学院学报》2010年第6期542-546,共5页Journal of Dalian Fisheries University

基　　金：国家"十一五"科技支撑项目(2006BAD09A01);国家"908"项目(908-01-ZH3);辽宁省教育厅高等学校科研计划项目(2008144);大连市科技攻关项目(2006B11NC109);大连海洋大学校人才引进项目(SYYJ200703)

摘　　要：应用杂交瘤技术制备了4株稳定分泌抗仿刺参Apostichopus japonicus体腔细胞单克隆抗体的杂交瘤细胞株,并对其单克隆抗体的特性进行了分析。结果表明:单抗1C7和1H7主要结合淋巴样细胞,单抗2E8可与所有体腔细胞结合,单抗4E9主要结合球形细胞;单抗2E8、4E9与仿刺参体腔液具有明显的交叉反应,单抗1C7和1H7则与体腔液无交叉反应;单抗1C7和4E9属于IgG1,2E8属于IgG2a,1H7属于IgM亚型;单抗1C7和1H7均可识别相对分子质量为37 000的蛋白,单抗2E8可同时识别相对分子质量为37 000和61 000两种蛋白,单抗4E9可识别相对分子质量为108 000的蛋白。该单抗的制备为进一步研究仿刺参体腔细胞和体腔液之间的关系,体腔细胞的分类、分布、发生、分化及功能提供了新的工具。Four monoclonal antibodies（MAbs）（1C7,1H7,2E8,and 4E9） against coelomocytes of sea cucumber Apostichopus japonicus were prepared by immunising BALB/c mice,and the characteristics were studied by the indirect immunofluorescence assay test（IIFAT）,indirect enzyme-linked immunosorbent assay（ELISA） and western-blotting.The MAb 1C7 and 1H7 were showed to have specificity for lymphoid cells,and MAb 4E9 for spherulocytes.The band MAb 2E8 was bound to all of coelomocytes.The MAb 2E8 and 4E9 were showed to be reacted with coelomic fluid components of the sea cucumber,and to be negative to MAb 1C7 and 1H7.The MAb 1C7 and 4E9 were IgG1 type,2E8 IgG2a type,and 1H7 IgM type.The western-blotting showed that MAb 1C7 and 1H7 recognised an antigen of molecular weight 37 000 protein.The MAb 4E9 108 000 protein,and MAb 2E8 reacted antigens 37 000 and 61 000 protein.This study is the preliminary step,and we will use the MAbs in study of the relationship between coelomocytes and coelomic fluid,and type,distribution,ontogenesis,differentiation and function of coelomocytes in the future.

关 键 词：仿刺参 体腔细胞 单克隆抗体 

分 类 号：Q813.2[生物学—生物工程]