【摘要】 目的筛选次黄嘌呤-氨基喋呤-胸腺嘧啶(HAT)敏感型产抗苏丹红Ⅰ单克隆抗体杂交瘤细胞株,为双特异性抗体的制备作好准备。方法在培养瓶中培养产抗苏丹红I单克隆抗体杂交瘤细胞株H6,取长到对数生长期的细胞,加入1.25μg/ml的8-氮鸟嘌呤,继续培养,当细胞长到60%左右时,弃去一部分细胞,加入2.5μg/ml的8-氮鸟嘌呤,继续培养,重复以上操作,倍比提高8-氮鸟嘌呤的含量,直至8-氮鸟嘌呤浓度达到20μg/ml,取细胞上清ELISA检测其产抗体情况。结果成功诱导筛选到HAT敏感的次黄嘌呤-鸟嘌呤磷酸核糖转移酶(HGPRT)缺陷型杂交瘤细胞6株,均能分泌抗苏丹红Ⅰ单克隆抗体。结论该方法能成功筛选HAT敏感型抗苏丹红Ⅰ单克隆抗体杂交瘤细胞。更多还原

【Abstract】 ObjectiveTo screen hypoxanthine aminopterin and thymidine(HAT) sensitive cell lines of Sudan red Ⅰ monoclonal antibody hybridoma bispecific antibody preparation. MethodsCulture Sudan red Ⅰ monoclonal antibody hybridoma cell line H6,add 1.25 μg / ml 8 azaguanine to the culture medium,continue to develop.When cells grow to about 60%,culture with 2.5 μg / ml 8 azaguanine,continue to develop.Repeat the above operation until the concentration of 8 azaguanine reached 20 μg / ml,and then ELISA detect whether it can produce antibody. ResultsSix cell strains of the HAT sensitive hypoxanthine guanine phosphoribosyl transferase(HGPRT) defective hybridoma were successfully got,which all secret anti Sudan red Ⅰ monoclonal antibody. Conclusion The method can successfully screen HAT sensitive and anti sudan Ⅰ monoclonal antibody hybridoma cell.更多还原