作　　者： ; ; ; ;

机构地区： 华南理工大学生物科学与工程学院

出　　处： 《华南理工大学学报（自然科学版）》 2008年第12期133-137,共5页

摘　　要： 为了获得镉螯合物特异性单克隆抗体,以异硫氰酸苄基乙二胺四乙酸为双功能螯合剂螯合镉离子制得半抗原,再分别与钥孔戚血蓝素或牛血清白蛋白偶联制得免疫原与包被抗原.经过抗原鉴定,免疫Balb/c小鼠,取小鼠的脾细胞与SP2/0骨髓瘤细胞融合制备杂交瘤,经筛选和2次克隆化等步骤,从制备的腹水中获取单克隆抗体.酶联免疫吸附测定结果表明,单克隆抗体3A9D9G7的腹水效价为4×10-4,单抗亚类为IgM,轻链为κ型,亲和常数为1.44×1010mol/L,灵敏度低于50μg/L,与其他金属螯合物的交叉反应率低于0.90%,说明该单克隆抗体能特异性识别镉螯合物,可用于环境样品与食品中残留镉的快速检测. In order to obtain the specific monoclonal antibody against cadmium chelate, 1-（4-isothiocyanobenzyl） ethylenediamine-N, N, N′, N′-tetraacetic acid （ITCBE） was used as the bifunctional chelator to prepare the hapten. The product was then coupled with keyhole limpet hemocyanin Cd-ITCBE-KLH or bovine serum albumin （BSA） to produce immunogen and coating antigen. After an identification of the antigen and the immunization of Balb/c mice, the hybridomas were screened via the fusion of spleen cells and SP2/0 myeloma cells, and positive clones against Cd-ITCBE-BSA were selected by subcloning. Finally, mAb 3A9D9G7 was obtained from the ascites. ELISA results indicate that mAb 3A9D9G7 with a κ-type light chain, whose ascites titer and apparent affinity constant are respectively 4 × 10 ^-4 and 1.44 × 10^10 mol/L, belongs to IgM subclass. It is of a sensitivity of lower than 50μg/L and a cross-reactivity rate of less than 0. 90%. Thus, mAb 3A9D9G7 is highly specific against cadmium chelate and is suitable for the rapid detection of cadmium residue in environment and food samoles.

关 键 词： 镉 双功能螯合剂 单克隆抗体 制备 鉴定

领　　域： [生物学]