抗肉毒神经毒素A(BoNT/A)单克隆抗体的制备与鉴定(英文)  

Generation and characterization of monoclonal antibodies against botulinum neurotoxin type A (BoNT/A)

作　　者：王景林[1] 康琳[1] 贾宏丽[1] 

机构地区：[1]军事医学科学院微生物流行病研究所全军微生物检测研究中心,北京100071

出　　处：《细胞与分子免疫学杂志》2004年第1期83-85,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：Supportedbythenationalhightechnologyresearchanddevelop mentprogram (863)ofchina (No.30 370 0 81 )andthenationalNaturalScienceFoundation

摘　　要：目的 :制备抗肉毒毒素A(BoNT/A)的单克隆抗体 (mAb)。方法 :用纯化的重组BoNT/A Hc片段免疫BALB/c小鼠 ,取其脾细胞与骨髓瘤Sp2 /0融合 ,经间接ELISA筛选和克隆化制备杂交瘤细胞系 ,及Western免疫印迹分析等方法对mAb进行特异性鉴定。结果 :获得 3株杂交瘤细胞株 :命名为4A8、2F7和 4F2 ,IgG亚类鉴定均为IgG1,腹水mAb的效价在 1× 10 -4～ 1× 10 -6之间。其中 ,4A8和 4F2能稳定分泌抗BoNT AmAb,并可特异性地识别重组BoNT/A和天然BoNT/A ,特别是 4A8可保护小鼠抵抗 10LD50 BoNT/A的攻击。结论 :成功地制备 3株特异性抗BoNT/AmAb ,并有 1株属于中和性mAb 。AIM: To generate the monoclonal antibodies (mAbs) against botulinum neurotoxin type A (BoNT/A). METHODS: BALB/c mice were intraperitoneally immunized with purified BoNT/A-Hc, and the splenocytes of immunized mice were fused with myeloma cells Sp2/0. Hybridoma cells were screened by indirect ELISA and monoclonal hybridoma cells was obtained using limited dilution. RESULTS: Three hybridomas, named 4A8, 2F7 and 4F2, producing the mAbs against BoNT/A, were successfully established, and the titer of ascitic mAbs ranged from 1×10 -4 to 1×10 -6. Identification of subclass showed that all the produced mAbs belonged to IgG1. 4A8 and 4F2 were stable in secreting anti-BoNT/A mAbs through three-month continuous culture and showed high specificity to recombinant BoNT/A-Hc and native BoNT/A. CONCLUSION: Anti-BoNT/A mAbs we generated have high specificity, which laid the foundation for the immunological detection of BoNT/A and clinical treatment of botulism in the future.

关 键 词：肉毒神经毒素A 肉毒杆菌毒素 单克隆抗体 

分 类 号：R971.92[医药卫生—药品]