副结核分枝杆菌McAb的某些特性鉴定

Partial Characterization Of McAbs To Mycobacterium Paratuberculosis

以超声波粉碎的副结核分枝杆菌地力株C_2抗原免疫Balb/c小鼠,经与NS—1骨髓瘤细胞三次融合,获得了3株能稳定分泌抗副结核分枝McAb的杂交瘤细胞系,其染色体数目为78～110条,分别为IgM和IgG_2。应用间接ELISA法测定细胞培养液、腹水和血清效价为10^(-2)～10^(-7)。交叉试验证明,McAbO_1与受试的各株副结核分枝杆菌反应强烈,与牛分枝杆菌和草分枝杆菌有微弱的交叉反应,与其他受试的分枝仟菌不反应,说明McAbO_1所对应的抗原是副结核分枝杆菌种特异性抗原之一。SDS—PAGE、免疫转印试验结果表明,McAb_1、O_2和O_3所对应的抗原分子量分别为38.5,55.5和6.5KD。

Three hybridomas which stably secrete antibodies to Mycobacterium paratuberculosis were obtained by three fusions of NS- 1 myeloma cells with spleen cells of mice immunized with M. paratuberculosis C2 culture sonicated. The chromosome number of hybridomas cells were between 78-110. As to the Ig subclass, they belongs to IgM and IgG2. The indirect ELISA liters of those McAbs, cultural supernatants ascites and sera are between 10-2 - 10-7. An antibody, designated McAbs strongly releted with different strains of M. paratuberculosis used in the test and faintly with the M. bovis and the M. phlei and not with the reference strains of the other mycobacteria tested. The result suggested the antigen of McAbs be one ef the mycobacteria specificity. The antigen molecular weight of the McAbO1 and O2 were 38. 5, 55.5 and 6. 5 KD by the SDS-PAGE and immunoblots assay.