抗同型磺基丙氨酸单克隆抗体的制备和定性  

Preparation and Characterization of Monoclonal Antibody to L-Homocysteic Acid

作　　者：刘长金[1] 李爱[1] 胡新武[1] 张亮品[1] 

机构地区：[1]华中科技大学同济医学院基础医学院生理学教研室,武汉430030

出　　处：《同济医科大学学报》2001年第5期410-412,共3页Acta Universitatis Medicinae Tongji

基　　金：国家自然科学基金资助项目 ( No. 3 880 3 11)

摘　　要：用戊二醛将 L-同型磺基丙氨酸 ( L- HCA)连接到牛血清白蛋白 ( BSA) ,形成 L- HCA- BSA连接物。用 L- HCA-BSA免疫小鼠后 ,收集脾淋巴细胞 ,并与骨髓瘤细胞融合。用含次黄嘌呤、氨基蝶呤和胸腺嘧啶核甙 ( HAT)培养基保留杂交细胞。用 EL ISA法筛选能够分泌与 L- HCA- BSA产生免疫反应的杂交细胞。通过 EL ISA抗体系列稀释试验和抗体吸收试验 ,确定抗体特异性。结果表明 ,抗 L- HCA单克隆抗体 ( Mc Ab) 2 D4能特异地与 L- HCA- BSA产生免疫反应。用 L- HCA- BSA吸收 L- HCA Mc Ab,可明显降低或取消 L- HCA抗体与 L- HCA- BSA的免疫反应。结果提示 :L- HCAMc Ab对 L- HCA- BSA具有特异性 ,可用于免疫组织和细胞化学研究 L- HCA在神经系统中的分布和定位。L-homocysteic acid (L-HCA) was conjugated to bovine serum albumin (BSA) with glutaral-ldehyde to form L-HCA-BSA conjugates. Monoclonal antibody (McAb) to L-HCA was developed following immunization with L-HCA-BSA conjugates, cell fusion, selection of hybridoma cells which produced antibody reacting with L-HCA-BSA with HAT (hypoxanthine, aminopterin, thymidine) medium. Serial dilution test and antibody absorption test on ELISA (enzyme-linked immunosorbent assay) were performed. The results indicated that McAb to L-HCA could react with L-HCA-BSA, but not with other amino acids-BSA conjugates so far tested. McAb absorbed with L-HCA-BSA conjugates could abolish or decrease immunoreactivity of L-HCA-BSA with McAb. It was suggested that McAb to L-HCA was specific for L-HCA-BSA and could be used for immunohistocytochemical localization and distribution of L-HCA in the nervous system.

关 键 词：L-同型磺基丙氨酸 单克隆抗体 酶联免疫吸附测定 制备 

分 类 号：R392－33[医药卫生—免疫学] R446.61[医药卫生—基础医学]