次黄嘌呤-鸟嘌呤磷酸核糖转移酶缺陷型产抗苏丹红Ⅰ单克隆抗体杂交瘤细胞的筛选  

Screening of Hypoxanthine Aminopterin and Thymidine(HAT) Sensitive and Anti Sudan Red Ⅰ Monoclonal Antibody Hybridoma Cell

作　　者：裘雪梅[1] 刘仁荣[1] 徐玲[1] 朱立鑫[1] 

机构地区：[1]江西科技师范学院生命科学学院,江西南昌330013

出　　处：《时珍国医国药》2012年第11期2669-2670,共2页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.30860240);江西省科技厅科技项目(No.2010GZN0022)

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。Objective To screen hypoxanthine aminopterin and thymidine （ HAT ） sensitive cell lines of Sudan red I monoclonal antibody hybridoma bispecific antibody preparation. Methods Culture 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. Results Six cell strains of the HAT sensitive hypoxanthine guanine phos- phoribosyl 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.

关 键 词：8-氮鸟嘌呤 次黄嘌呤-氨基喋呤-胸腺嘧啶(HAT) 杂交瘤 

分 类 号：R392.4[医药卫生—免疫学]