分泌抗鼠疫耶尔森菌F1抗原单克隆抗体杂交瘤株的建立  

Establishment of hybridoma cell strains secreting McAbs against Yersinia Pestis F1 antigen

作　　者：尹家祥[1] 陈平[1] 杜春红[1] 董兴齐[1] 于国林[1] 

机构地区：[1]云南省地方病防治所,大理671000

出　　处：《中国地方病学杂志》2005年第5期566-568,共3页Chinese Jouranl of Endemiology

基　　金：国家自然科学基金资助项目(30160078);云南省科技攻关计划资助项目(2002NG17)

摘　　要：目的应用杂交瘤技术,制备分泌抗鼠疫F1抗原单克隆抗体的杂交瘤细胞株。方法用鼠疫菌F1抗原免疫雌性BALB/c小鼠,检测小鼠血清中的F1抗体,选抗体滴度最高的小鼠用于细胞融合,融合前3天F1抗原经腹腔注射加强免疫1次,然后取其脾脏细胞与处于对数生长期的SP2/0骨髓瘤细胞混合,50%的PEG作为细胞融合剂,HAT培养液选择培养融合后的细胞,间接ELISA法检测细胞培养上清中的F1抗体,阳性杂交瘤细胞用有限稀释法进行克隆及再克隆。结果获得共计100余株分泌F1单克隆抗体的杂交瘤细胞株,3株进行了4次克隆化,经8个月保存后抗体分泌稳定。结论应用杂交瘤技术,在云南首次成功制备了3株能稳定分泌抗鼠疫杆菌F1抗原的单克隆抗体杂交瘤细胞株,建立了杂交瘤技术的实验程序。Objective To establish hybridoma cell strains secreting FI monoclonal antibodies （McAbs） against Yersinia pestis（ Y. pestis ） F1 antigen using hybridoma technology, Methods Healthy female BALB/c mice were immunized with F1 antigen, Blood was collected from the mice via caudal vein, and the sera were tested for anti-F1 antibody titres by indirect enzyme-linked immunosorbent assay （ELISA）, The mouse with the highest antibody titre was again injected with F1 antigen into abdominal cavity three days before cell fusion to enhanced the immunity. The spleen cells of the immunized mouse were fused with myeloma cells SP2/O placed in the logarithmic growth phase using 50% polyethylene glycol （PEG） as cell fusion reagent, Hybridoma cells were selected on hypoxanthine aminopterin and thymidine （HAT） medium, The supernatants of hybridoma cells were tested using indirect ELISA for the presence of F1 antibodies. Limited dilution was used to clone the positive hybridoma cells that produced F1 McAbs, Results More than 100 hybridoma cell strains that could secret F1 McAbs were obtained, 3 strains stably secreting F1 McAbs were obtained after four clonings, Conclusion Using hybridoma technology, 3 hybridoma cell strains that can stably produce F1 McAbs against Y, pestis F1 antigen have been firstly successfully established in Yunnan, Experimental procedures of hybridoma technique have been set up.

关 键 词：耶尔森菌 鼠疫 F1抗原 杂交瘤细胞 单克隆抗体 

分 类 号：R392[医药卫生—免疫学]