大豆凝集素单克隆抗体的制备及鉴定  

Preparation and Identification of Monoclonal Antibody against Soybean Agglutinin

作　　者：张海全[1,2] 车东升[1,2] 刘飞飞[1,2] 穆成龙[1,2] 谷琳琳[1,2] 孙泽威[1] 秦贵信[1] 

机构地区：[1]吉林农业大学,动物生产及产品质量安全教育部重点实验室,吉林长春130118 [2]吉林农业大学动物科学技术学院,吉林长春130118

出　　处：《中国畜牧兽医》2012年第9期79-82,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(30871802)

摘　　要：为获得分泌抗大豆凝集素(SBA)单克隆抗体的杂交瘤细胞,以纯化的SBA为抗原,免疫BALB/c小鼠,加强免疫3d后取小鼠脾细胞与骨髓瘤细胞(SP2/0)融合,应用有限稀释法和间接ELISA方法克隆筛选出2株分泌抗SBA单克隆抗体的杂交瘤细胞系A7、F12。细胞上清抗体效价均在1∶2×103以上,腹水抗体效价均为1∶1×106,单抗亚型鉴定结果均为IgG2b型,分子质量为189.6ku,亲和常数为7.1×107 mol/L,Western blotting结果表明,2株单抗具有较高的特异性。该McAb的制备为建立SBA定量检测方法奠定了基础。To prepare hybridoma cells secreting specific monoclonal antibody against soybean agglutinin（SBA）,the BALB/c mice were immunized by the purified SBA.The spleen cells isolated from the mice in the highest titer were fused with SP2/0 myeloma cells after three days of last immunization.Two hybridoma cell lines secreting McAb against SBA were establishedafter screening by ELISA and cloning four times by limiting dilution,named as A7,F12.The titers of secreting antibody from cell culture supernatants were both above 1∶2×10^3,while above 1∶1×10^6 from ascites of the mice.The subtypes of the monoclonal antibodies were both IgG2b,the molecular weight was 189.6ku,the affinity constant was 7.1×10^7 mol/L,Western blotting showed high specificity with SBA.The preparation of the McAb against SBA provided an useful tool for quantitative determination of SBA.

关 键 词：大豆凝集素 单克隆抗体 制备 鉴定 

分 类 号：Q78[生物学—分子生物学]