抗新型隐球菌荚膜相关蛋白CAP10单克隆抗体的制备及鉴定

Preparation and Evaluation of Monoclonal Antibodies Specific to CAP10 Protein of Cryptococcus neoformans

目的:以新型隐球菌荚膜相关蛋白CAP10为靶抗原,制备并鉴定特异性抗CAP10的单克隆抗体。方法:用纯化的重组CAP10免疫BALB/c小鼠,血清抗体效价达到适当水平时进行细胞融合;经多次亚克隆筛选出分泌特异性抗体的细胞株,制备单抗腹水并进行抗体效价测定及亚类鉴定。结果:获得11株能稳定分泌抗新型隐球菌荚膜相关蛋白CAP10的单克隆抗体杂交瘤细胞株,抗体效价高且抗原特异性强。结论:获得了针对新型隐球菌荚膜相关蛋白CAP10的单克隆抗体,为深入研究CAP10蛋白的功能,以及临床新型隐球菌的检测和血清型分析奠定了基础。

Objective： To produce monoclonal antibodies（mAb） specific to CAPIO protein of Cryptococcus neofor roans. Methods： The BALB/c mice were immunized with the recombinant CAP10-1 （70-250 aa）, and fusion of lymphocyte and myeloma Sp2/0 cell was performed by conventional method. Antibody titers and specificities of mAb were measured by ELISA followed production of mAb. Results： Eleven strains of hybridoma cell lines with the stable secretion of mAb to protein CAP10 were generated, and all the mAb only reacted with the recombinant CAP10-1. Conclusion： High titers of the specific mAb to the conserved fragment of CAPIO of C.neoformans were successfully produced, which is helpful for further study of the function of CAP10 and clinical diagnosis and sero- type analysis for infections of C.neoformans.