抗莱克多巴胺单克隆抗体杂交瘤细胞株的建立及其免疫学特性鉴定

Establishment of Hybridoma Cell Lines Secreting Anti-ractopamine Monoclonal Antibody and Identification of Their Immunological Properties

莱克多巴胺(RAC)经化学修饰引入功能基团羧基,合成半抗原RAC-戊二酸酐半醛化合物(RAC-SA),用混合酸酐法(MA)将RAC-SA偶联于牛血清白蛋白(BSA),合成人工抗原BSA-RAC,用红外(IR)、紫外(UV)和凝胶电泳(SDS-PAGE)对其进行鉴定;用BSA-RAC免疫Balb/C小鼠,细胞融合技术建立抗RAC的单克隆抗体(RAC mAb)杂交瘤细胞株,体内诱生腹水法制备RAC mAb,并鉴定其免疫学特性。结果表明,BSA-RAC偶联成功,偶联率为24.5:1;筛选出3F10、3H12、4D8共3株杂交瘤细胞,其中最好的4D8株间接ELISA效价细胞培养上清为1:1.28×103,腹水为1:6.4×105,同种型为IgG1/κ,亲和常数(Ka)为1.65×1010L/mol,对RAC的半数抑制浓度(IC50)为5.7ng/ml,与多巴酚丁胺的交叉反应率(CR)为22.3%,与其他化合物无CR。

The active group carboxyl was introduced to ractopamine(RAC) and formed RAC semialdehyde(RAC-SA) which has hapten structure by chemical modification.The method of mixed anhydride(MA) was used to conjugate RAC-SA to bovine serum albumin(BSA) and obtain artificial antigen BSA-RAC identified by infrared(IR),ultraviolet(UV) and SDS-PAGE.Balb/C mice were immunized with BSA-RAC and hybridoma lines that secrete monoclonal antibody against RAC(RAC mAb) were established through cell fusion and then screened using in...