鸡毒支原体烯醇化酶单克隆抗体研制及黏附阻断

Preparation and adherence inhibition by monoclonal antibodies to enolase of Mycoplasma gallisepticum

通过对鸡毒支原体烯醇化酶的克隆和原核表达,利用表达产物免疫小鼠,制备单克隆抗体,以超声波裂解的MG全菌作为筛选抗原,结果获得了6株特异性单抗,分别命名为:1A4、2D7、2E4、2F4、3B4和3D8。经间接ELISA、免疫荧光试验及Western blot检测发现:1A4具有荧光效价和高滴度的ELISA效价,但无免疫印迹效价;而其他5株单抗,免疫印迹呈阳性,但无表面染色的活细胞免疫荧光。在所有6株单抗中,仅1A4单抗有MG细胞黏附抑制作用,可导致MG的黏附率降低34.40%,而且1A4与其他5种单抗混合后,则黏附率可下降57.69%。这些抗体的制备将为烯醇化酶生物学活性研究奠定基础。

To prepare the monoclonal antibodies against MG enolase, we cloned the enolase of Mycoplasma gallisepti- cure ,and immunized the mice with the production of the prokaryotic expression,and used the supersonic lytic MG as screening antigen. At last, 6 specific monoclonal antibodies were obtained, named 1 A4,2D7,2E4,2F4,3B4 and 3D8. Screening by indirect ELISA,immunofluorescence assy （IFA）,and Western blot analysis, 1A4 had fluorescence and high ELISA titer,while it didn＇t have immunoblotting titer. However,other 5 mAbs had positive immunoblotting ti- ter, while the surface stained cells had no positive results by IFA. The obtained mAbs had a satisfactory immunoreac- tivity. The preparation of these antibodies established the foundation of the further research of the biological func- tions of the MG enolase.