抗青霉素单克隆抗体的制备及初步应用.doc

抗青霉素单克隆抗体的制备及初步应用    Preparationandprimaryapplicationofmonoclonalantibodyagainstbenzylpenicillin     [Abstract]AIM:Todevelopmonoclonalantibody(mAb):Penicillin,asakindofhapten,pleteantigenandthenwasusedtoimmunizeBALB/?:NinehybridomacellsstablysecrectingmAbwerepreparedthroughcellfusion,screeningandcloning,?antibodySandwichELISAforrelativequantitativeanalysisofbenzylpenicilloylproteinwasestablishedwithasensitivityof870U/%andtheaverageintra?andinter?coefficientofvaritation(CV)%%:?antibodySandwichELISAforrelativequantitativeanalysisofbenzylpenicilloylproteinwasestablished.     [Keywords]benzylpenicillin;benzylpenicilloyl;mAb;double?antibodySandwichELISA     [摘要]目的:制备抗青霉素的单克隆抗体(mAb)并建立双抗体夹心ELISA检测方法,对临床上引起青霉素过敏反应的过敏原青霉噻唑蛋白进行研究。方法:将半抗原青霉素和载体蛋白偶联后免疫BALB/c小鼠,应用杂交瘤技术建立稳定分泌抗青霉素mAb的杂交瘤细胞株。常规制备腹水,用辛酸?硫酸铵法纯化,并对纯化的mAb进行特异性鉴定。通过对不同抗体组合的分析和条件的优化,建立检测过敏原的双抗体夹心ELISA方法。结果:经细胞融合、筛选及克隆化,共获得9株稳定分泌抗青霉