大豆凝集素单克隆抗体的制备及鉴定

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3 d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189.6 ku,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。

Preparation and Identification of Monoclonal Antibody against Soybean Agglutinin

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∶2×103,while above 1∶1×106 from ascites of the mice.The subtypes of the monoclonal antibodies were both IgG2b,the molecular weight was 189.6ku,the affinity constant was 7.1×107 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.