腐马素B_1单克隆抗体的制备与鉴定  

Preparation and Identification of Monoclonal Antibodies Against Fumonisin B_1

作　　者：钟秋芳[1] 肖希龙[1] 章红[1] William P.Norred F.Meredith 

机构地区：[1]中国农业大学,北京100094 [2]美国农业部毒理与真菌毒素研究中心

出　　处：《畜牧兽医学报》2002年第3期267-270,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家"九五"攻关专题"饲料添加剂质量检测技术研究"资助

摘　　要：本研究采用戊二醛一步法制备免疫抗原FB1 KLH和包被抗原FB1 BSA ,免疫BALB/c小鼠 ,建立间接酶联免疫吸附法 (I ELISA)并测定血清效价。结果表明 :FB1 BSA最适包被浓度为 1μg/ml,抗体最适稀释度为 1∶10 0 0 0 ,血清效价可达 1∶12 80 0 0。第 4次免疫后 ,通过淋巴细胞杂交瘤技术建立分泌FB1单克隆抗体的杂交瘤细胞株 ,融合率为 96 % ,阳性率为 98%。细胞上清抗体ELISA效价为 1∶3 2× 10 6,腹水抗体效价为 1∶8× 10 8。经鉴定 ,该株单抗的亚类为IgG1,分子量为 185 4KDa ,染色体数目介于 96～ 10 4之间 ,亲和常数为 1 16× 10 7M-1,与另三种常见的真菌毒素 (串珠镰刀菌素 ,T 2毒素 ,玉米赤霉烯酮 )和两种载体蛋白 (牛血清白蛋白 ,血蓝蛋白 )无交叉反应 。In this study,one hybridoma was obtained against FB 1,the most abundant in Fumonisins.The results were as follows:FB 1 KLH using as immunogen and FB 1 BSA using as solid phase antigen were prepared with one step glutaraldehyde method.BALB/c mice were immunized with FB 1 KLH and an indirect ELISA was developed.The result showed:the optimal concentration of coating antigen was 1 μg/ml,the concentration of antibody was 1∶ 10 000 and the titer could reach 1∶ 128 000 .The fusion was performed after the forth immunization with hybridoma technique.The fusion rate was 96%,the positive rate was 98%.The titer of supernatant and that of ascitic fluid were 1∶3 2×10 6 and 1∶8×10 7 respectively.The characterization of antibody was identified,the results indicated:the subclass was IgG 1,the molecular weight was 185.4KDa,the chromosomal numbers were between 96～104,the affinity constant was 1.16×10 7M -1 .No cross reactivity was seen between the antibody and the other three mycotoxins (Moniliformin,T 2 toxin,Zearalenone) as well as the two carrier proteins (BSA,KLH),the antibody had excellent stability.

关 键 词：腐马素B1 单克隆抗体 制备 鉴定 

分 类 号：S852.66[农业科学—基础兽医学] S852.4[农业科学—兽医学]