抗金葡菌多肽单克隆抗体制备及初步鉴定

目的:研制抗金葡菌多肽单克隆抗体(mA b)。方法:抗金葡菌多肽(PH-SA)抗原采用脾内注射、醋酸纤维膜皮下包埋、完全佐剂3种方法免疫Balb/C小鼠,常规方法细胞融合,用EL ISA间接法筛选阳性细胞并测定相对亲和力,双抗体竞争试验和双抗体竞争抑制试验识别PH-SA表位。结果:获4株可分泌PH-SA mA b的杂交瘤细胞株。免疫球蛋白亚类测定均为小鼠IgG1亚类,相对亲和力依次为G11>B9>H3,4株mA b可能抗相同抗原表位。结论:获4株PH-SA mA b的杂交瘤细胞株为PH-SA的鉴定提供了可能性。

Preparation and preliminary characterization of monoclonalantibodies against Pheromonicin-and-Staphylococcus aureus

Objective: To obtain monoclonal antibodies(mAbs) against Pheromonicin-and-Staphylococcus aureus(PH-SA).Methods: BALB/c mice were immunized by immunogen PH-SA in three different ways-intrasplenic inoculation,subcutaneous cellulose acetate membrane embedding,and complete Freud's adjuvants.Splenocytes of immunized mice and Sp2/0 cells were fused conventionally,and indirect ELISA was applied to screen for positive mAb-secreting hybridoma cells and detect mAb relative affinity.The epitopes on PH-SA recognized were analyzed by double antibody competition test and double antibody competition inhibition test.Results: Four hybridoma cell lines secreting mAbs against PH-SA,all of IgG_1 subclass,were obtained,and the relative affinity lies in G_(11)>B_9>H_3.the antibodies might recognize the same epitope on PH-SA.Conclusion: The four obtained hybridoma cell lines secreting mAbs against PH-SA provide the possibility of PH-SA identification.