Objective To develop monoclonal antibodies specific for infectious vaginosis (bacterial vaginosis, trichomonas vagi-nalis and fungal vaginitis). Methods The genes of pathogens in infectious vaginosis were analyzed and gene sequences, encoding specific proteins, were amplified to construct recombinant antigen AP33. Then Aalb/c mice were immunized by AP33 and specific monoclonal antibodies were prepared by hybridoma technology. The tilter,subtype and the light chain of acquired antibodies were analyzed by indirect enzyme linked immunosorbent assay,and the concentration, purity and molecular weight of purified antibodies were analyzed by bicinchoninic acid(BCA) method and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Results The purity of monoclonal antibody,produced by hybridoma cell line,reached 90% ,with concentration of 2. 41 -5. 68 mg/ mL and titers of 1 : 105 -1 : 107. Produced antibody was IgGl and with light chain of /c-type. Conclusion Monoclonal antibodies, specific for infectious vaginosis,were successfully produced,with high purity and specificity,could be used for pharmacology, diagnostics and related scientific researches,and showed tremendous marketing potential in future.%目的 研发感染性阴道疾病(细菌性阴道炎、滴虫性阴道炎和霉菌性阴道炎)的特异性单克隆抗体.方法 分析感染性阴道疾病病原体的基因谱,扩增特异蛋白的基因序列,基因工程重组抗原AP33,免疫Balb/c 小鼠,运用杂交瘤技术得到特异性单克隆抗体;采用间接酶联免疫吸附试验(ELISA)进行抗血清效价测定及抗体的亚类及轻链型鉴定;采用BCA法、聚丙烯酰胺凝胶电泳(SDS-PAGE)法分析纯品抗体的蛋白浓度、纯度和相对分子质量.结果 杂交瘤细胞株产生的单克隆抗体纯度均达到了90%以上,抗体浓度2.41～5.68 mg/mL,抗体效价在1∶105～1∶107之间;抗体类别均为IgG1,κ型.结论 本研究获得了3种感染性阴道疾病纯度高、特异性强的单克隆抗体,将为药理学、诊断学及相关学科提供研究材料,具有广阔的市场前景和巨大的经济效益.
展开▼
