CAJ | 학술논문

为制备绵羊早孕因子( EPF)单克隆抗体,以纯化的EPF重组蛋白为抗原免疫BALB/c小鼠,免疫4次后,取小鼠的脾细胞与骨髓瘤细胞SP2/0融合,采用间接ELISA方法筛选单克隆抗体细胞株,将筛选的细胞株注入小鼠腹腔得到腹水型抗体,用G-sepharose柱亲合层析法对小鼠腹水型抗体进行纯化,应用SDS-PAGE和Western blot分析抗体纯化后的纯度及特异性。结果显示,细胞融合后得到了1株抗绵羊EPF单克隆抗体的细胞株B4 D5,纯化后的腹水型单克隆抗体纯度较高,具有较强的特异性,应用单克隆抗体检测绵羊妊娠准确率为81.3%。表明,制备的绵羊早孕因子单克隆抗体为特异性抗绵羊EPF抗体。
위제비면양조잉인자( EPF)단극륭항체,이순화적EPF중조단백위항원면역BALB/c소서,면역4차후,취소서적비세포여골수류세포SP2/0융합,채용간접ELISA방법사선단극륭항체세포주,장사선적세포주주입소서복강득도복수형항체,용G-sepharose주친합층석법대소서복수형항체진행순화,응용SDS-PAGE화Western blot분석항체순화후적순도급특이성。결과현시,세포융합후득도료1주항면양EPF단극륭항체적세포주B4 D5,순화후적복수형단극륭항체순도교고,구유교강적특이성,응용단극륭항체검측면양임신준학솔위81.3%。표명,제비적면양조잉인자단극륭항체위특이성항면양EPF항체。
Inorder to preparate anti-sheep early pregnancy factor ( EPF ) monoclonal antibodies and identify their specificities,spleen cells from BALB/c mice immunized four times with purified EPF were fused with homology myeloma cells( SP2/0 ) . The cell strains of monoclonal antibodies were detected by the indirect ELISA method, and ascitic fluid were obtained by injecting cell strains to enterocoelia of mice. Monoclonal antibodies were purified by G-sepharose column affinity chromatography. Purity and spe-cificity of monoclonal antibodies after purification were detected by SDS-PAGE and Western blot. A cell strain of anti-sheep EPF monoclonal antibodies was obtained after the cell fused ( B4 D5 ) . Anti-sheep EPF monoclonal antibodies were obtained with a high purity and strong specificity after purification,and preci-sion rate of pregnancy was 81. 3%. The monoclonal antibodies had a specificity EPF in sheep.