抗人IgG和抗人IgM单克隆抗体的特性鉴定及应用

Characterization and Application of Anti-Human IgG and Anti-Human IgM Monoclonal Antibodies

目的:制备特异性高的抗人Ig G和抗人Ig M单克隆抗体,并应用于临床血清学检测试剂盒,为其提供优质的二抗。方法:以人Ig G和Ig M为抗原,免疫BALB/c小鼠,通过传统杂交瘤融合和筛选技术制备抗人Ig G和抗人Ig M单克隆抗体;经体内诱生法制备腹水并纯化;采用酶联免疫吸附试验(ELISA)和Western印迹进行交叉筛选和特异性鉴定,并对筛选出的单抗进行辣根过氧化物酶(HRP)标记,再与市售的检测病原微生物抗体试剂盒中对应的HRP标记的二抗(抗人Ig G和抗人Ig M)做对比分析。结果:筛选到2株可稳定分泌抗人Ig G单抗和2株抗人Ig M单抗,经交叉反应、Western印迹及对比实验分析,证明这4株单抗效价高、特异性好,比临床试剂盒中的酶标二抗特异性强、敏感度高。结论:获得4株特异性强、敏感度高的抗人Ig G和抗人Ig M单克隆抗体,可为各种病原微生物血清学的检测提供二抗试剂,具有良好的应用价值。

Objective： To prepare the high-specificity anti-human IgG and anti-human IgM monoclonal antibodies （mAbs）, and to apply to clinical serological test kits, and to provide the high-quality secondary antibodies. Methods： BALB/c mice were immunized by the antigens of the human IgG and IgM. The anti-human IgG and anti-human IgM mAbs were prepared by traditional hybridoma fusion and screening techniques, the ascites was prepared by in vivo induction and purified, cross-screening and specificity identification by ELISA and Western blot assays. The selected mAbs were labeled with horseradish peroxidase（HRP）, then compare with the HRP-labeled secondary antibody of pathogenic microorganisms detection kit. Results： Two strains of anti-human IgG mAbs and two strains of anti-human IgM mAbs were screened. The results of crossing-reaction experiments, Western blot experiments and comparative experiments demonstrated that the four mAbs had high titer and specificity, and they were more specific and sensitive than the enzyme-labeled secondary antibodies in clinical kits. Conclusion： A total of 4 anti-human IgG and anti-human IgM mAbs with high specificity and sensitivity were obtained, which can provide secondary antibody reagents for serological detection of various pathogenic microorganisms.