1组曲霉单克隆抗体的制备与鉴定

Preparation and characterization of a panel of specific monoclonal antibodies to aspergillus

目的制备并鉴定一组抗曲霉不同抗原的单克隆抗体。方法采用烟曲霉细胞壁抗原成分、分泌抗原和灭活分生孢子,分别免疫BALB/c小鼠,制备单克隆抗体,免疫荧光法鉴定单克隆抗体与曲霉属和念珠菌属抗原的交叉反应。结果获得29株稳定分泌抗曲霉单抗的杂交瘤细胞株,其中用烟曲霉细胞壁抗原成分免疫获得11株,用分泌抗原免疫获得13株,用孢子免疫获得5株;Ig亚类鉴定,11个克隆株为IgG1亚类,3个克隆株为IgG3,15个克隆株为IgM。免疫荧光法鉴定29株单抗特异性识别烟曲霉细胞壁抗原,与其他曲霉抗原有交叉反应。结论29株单克隆抗体,对于建立侵袭性曲霉感染早期诊断方法、筛选曲霉保护性抗体以及研究抗体保护机制奠定了实验基础。

Objective To produce and characterize a panel of monoclonal antibodies（mAbs） specific to variable antigens of aspergillus. Methods BALB/c mice were immunized with antigenic components of aspergillus fumigatus （AF） cytoderm, excreting antigens and inactivated conidiospore respectively to produce mAbs, and the cross-reactions between these mAbs and aspergillus or candida antigens were characterized with immumofluorescence （IF） method. Results Twenty-nine strains of hybridoma cells excreting anti-aspergillus Abs stably were obtained, in which 11 strains were immunized with Cas, 13 strains with Eas, and 5 strains with spores. With IF method, all of 29 mAbs reacted with antigenic components of aspergillus fumigatus cytoderm specially, but not with aspergillus antigens of other species. Conclusion Twenty-nine mAbs specific to aspergillus antigens were characterized, which could lay a laboratory foundation to establish the early diagnostic method and screen the protective Ab of invasion aspergillus infections.