摘要
为进一步研究催产素分子的生理功能及空间结构 ,制备识别其特有表位的单抗。用改良的碳二亚胺二步法交联催产素和牛甲状腺球蛋白 ,用常规方法制备针对催产素分子的单抗 ,测定了抗体的亚类、亚型和相对亲和力 ,并采用竞争ELISA法 ,检测各株细胞分泌的单抗和几种催产素的拟似物的交叉反应程度。结果制备出了 3株分泌高效价抗体的杂交瘤细胞 4G11、 2B11和 3F1,其亚类均为IgG1,亚型均为κ型 ,相对亲和力均较高。 3株中 4G11和 2B11可以特异性地和催产素反应 ,而 3F1与AVP、VAS及ASU有较强的交叉反应。
To further study the physiological functions and cubic structure of Oxytocin,attempt to prepare monoclonal antibodies with special epitopes to this molecule had been made in the present study By using the innovated two steps carbodiimide method,Oxytocin was conjugated with bovine thyroglobulin and this conjugate was used as immunogen Hybrodoma cell lines were established by the routine method,and isotypes as well as the relative affinities of the antibodies identified By means of the competitive ELISA,the degrees of cross reactivities of monoclonal antibodies with the Oxytocin analogues AVP,VAS and ASU were analyzed Three strains of hybridoma cell lines 4G11,2B11 and 3F1 were obtained thereby,all of which secreted very high titers of specific antibodies,and all were of IgG1 for heavy chain and κ isotype for light chain,and all showed great relative affinities to Oxytocin The 4G11 and 2B11 antibodies reacted specifically with Oxytocin,but 3F1 cross reacted strongly with AVP,VAS and ASU These results demonstrate that monoclonal antibodies to Oxytocin can be used in the study to block the biological functions of Oxytocin
出处
《上海免疫学杂志》
CSCD
北大核心
2001年第3期180-181,共2页
Shanghai Journal of Immunology
基金
国家自然科学基金资助项目 (No 3 2 83 0 170 )
