抗结核杆菌单克隆抗体的研制及初步鉴定

Production and Partial Characterization of Monoclonal Antibodies to Mycobacterium Tuberculosis

运用单克隆抗体技术 ,以结核杆菌H37Rv的培养滤液免疫Balb/c小鼠 ,以H37Rv及BCG抗原同时筛选杂交瘤细胞 ,得到了 7株稳定分泌的抗结核杆菌的杂交瘤细胞株 ,并对它们的特性作了初步鉴定。在 7株单克隆抗体杂交瘤中 ,5株 (2C5、2C11、2D1、2E11及 3D3)为IgG1亚类 ,另两株 (3C3及 5D8)为IgM。腹水经ELISA检测 ,效价可达 10 -3 ～ 10 -4 ,同时应用ELISA及免疫印迹试验分析了 7株单抗的种间反应性 ,结果 7株单抗与其它结核杆菌有不同程度的交叉反应 ,其中 1株单抗 (5D8)与大肠杆菌超声抗原也有明显交叉反应 ,提示结核抗原组分十分复杂 ,不仅分支杆菌种间有共同抗原决定簇 ,甚至与其它种属也有相同或相似的抗原决定簇。将这

Seven monoclonal antibodies to Mycobcterium tuberculosis were generated by immunization of Balb/c mice with Mycobacterium tuberculosis H 37 Rv culture filtrate.In an attempt to select hybridomas producing Mycobaterium tuberculosis_specific immunoglobulins,culture supernatants were tested in ELISA against Mycobacterium tuberculosis and Mycobacterium bovis BCG antigens.Five MAbs designated as 2C 5,2C 11 ,2D 1,2E 11 and 3D 3 were IgG 1 subclass and the other two MAb designated as 3C 3 and 5D 8 was IgM class.The ELISA titres of the ascites were 10 - 3～10 - 4.Interspecies reactivity of the MAbs was analyzed by ELISA and immunoblots.Crossreactivity of these antibodies with other mycobacterial antigens was observed and one MAbs(designated as 5D 8) showed broad cross_reactivity even with E.coli sonicate.The results indicated the complexity of the components of the Mycobacterium tuberculosis antigens.The characterization of the MAbs and their applications to the serodiagnosis of tuberculosis need further evaluation