抗微囊藻毒素单克隆抗体胶体金标记物的制备及鉴定  

Production and characterization of gold labeled monoclonal antibody against MC-LR

作　　者：赵晓联[1] 孙蔚榕[2] 孙秀兰[1] 江涛[3] 张银志[1] 汤坚[1] 

机构地区：[1]江南大学食品学院,无锡214036 [2]江苏省微生物研究所有限公司所 [3]中国疾病预防控制中心营养与食品安全

出　　处：《卫生研究》2006年第4期442-445,共4页Journal of Hygiene Research

基　　金：国家"十五"科技专项(No.2001BA804A26);江苏省自然科学基金项目(No.BK2002406)

摘　　要：目的制备抗微囊藻毒素(MC-LR)单克隆抗体胶体金标记物,并利用光谱分析来探讨结合物的合成机理。方法用杂交瘤技术生产并纯化的抗MC-LR的单克隆抗体,利用胶体金标记技术合成20nm粒径的胶体金与抗MC-LR单克隆抗体的结合物,通过光谱分析对偶联物中单克隆抗体的构象进行分析,ELISA法检测偶联物中抗体的免疫性。结果抗MC-LR单克隆抗体的效价达108,IC50抑制浓度为3ng/ml,为IgG2a亚型,分子量为1·5×105,与MC-LW、MC-LF无明显交叉反应。胶体金与抗体偶联物的颗粒圆整,光谱分析结果显示抗体蛋白的特征官能团没有发生明显变化,与胶体金粒子作用后结构略加紧密。ELISA结果显示偶联物仍然保持免疫原性。结论抗MC-LR单克隆抗体胶体金标记偶联物中抗体蛋白结构和抗原决定簇的位点没有改变,仍具有免疫原性,偶联物的稳定性良好。Objective Production of the gold labeled monoclonal antibody against MC-LR, Study the simple mechanism of the conjugation 3y the spectral analysis, Methods Production and purification monoclonal antibodies against MC-LR by hybridoma technology, Production the 20nm colloidal gold and label the monoclonal antibody. Study the gold - McAb conjugate by the spectroanalysis. Results Monoclonal antibody produced by the hybridoma cells were tested for subtypes and designated as IgG2a for 8C11.The titers of antibody in ascites was 108.The IC50 for MC-LR was around 3ng/ml. The IgG molecule weight was 1.5×10^5 , The structure of the antibody was altered to more tighter and stabler, Conclusion Product the gold labeled monoclonalantibody against MC-LR conjugate successfully and keep the antigebinding determinant.

关 键 词：抗MC-LR单克隆抗体 胶体金 标记偶联物 制备鉴定 

分 类 号：R123.5[医药卫生—环境卫生学]