抗A型产气荚膜梭菌α毒素单克隆抗体杂交瘤细胞系的建立与中和效应  

Establishment of Hybridoma Cell Lines Secreting Monoclonal Antibodies against Alpha Toxin of Clostridium Perfringens Type A and the Netrulization Effect of the Produced McAbs

作　　者：许崇波[1,2] 马从林 朱平[1,2] 石瑾琪[1,2] 姚湘燕[1,2] 冯书章[1,2] 

机构地区：[1]解放军农牧大学军事兽医研究所 [2]山西省大同市第二卫生防疫站

出　　处：《卫生研究》1998年第S1期55-57,共3页Journal of Hygiene Research

摘　　要：用提取的Ａ型产气荚膜梭菌α毒素包涵体免疫ＢＡＬＢ／ｃ小鼠，取小鼠脾细胞与ＳＰ２／０骨髓瘤细胞进行融合和克隆化，经ＥＬＩＳＡ筛选，共获得７株稳定分泌单克隆抗体（ＭｃＡｂ）的杂交瘤细胞株，分别命名为１Ａ８、１Ｃ３、１Ｄ５、１Ｄ８、１Ｆ１、１Ｈ１和２Ｅ３。经鉴定，７株ＭｃＡｂ的Ｉｇ亚类有ＩｇＧ１（１Ｄ８）、ＩｇＧ３（１Ａ８、１Ｃ３和２Ｅ３）和ＩｇＭ（１Ｄ５、１Ｆ１和１Ｈ１）。细胞培养上清和腹水抗体效价分别为：１∶５１２～１∶１０２４和１∶１０６～１∶１０８。尤为重要的是，２Ｅ３杂交瘤细胞株分泌的ＭｃＡｂ不仅能够中和α毒素的磷脂酶Ｃ活性和溶血活性。おhe spleen cells from BALB/c mice immunized with purified inclusion bodies of alpha toxin of Clostridium perfringens were fused with SP2.0 myelomacells.Seven hybridoma cell lines which steadily secreted monoclonal antibodies were obtained by detecting with ELISA.These hybridoma cell lines were designated as 1A8 1C3 1D5 1D8 1F1 1H1 and 2E3 respectively,and their isotype subclass were IgG1(1D8) IgG3(1A8 1C3 and 2E3)and IgM(1D5 1F1 and 1H1).The ELISA titers of cell supernatant and ascites were 1∶512 to 1∶1024 and 1∶106 to 1∶108 respectively.Especially,monoclonal antibodies secreted by 2E3 hybridoma cell line could neutralize phospholipase C and the hemolytic activities of alphatoxin,and could provide protection for mice against the challenge of lethal dose of alpha toxin of Clostridium perfringens type A.

关 键 词：A型产气荚膜梭菌 Α毒素 单克隆抗体 中和效应 

分 类 号：Q933[生物学—微生物学]