抗人心肌肌钙蛋白T杂交瘤细胞系的建立及初步鉴定  

Establishment and Identification of Hybridoma Cell Lines Secreting Anti-human Myocardial Troponin T Antibody

作　　者：李跃辉[1] 周国华[1] 胡锦跃[1] 谢平丽[1] 李官成[1] 

机构地区：[1]中南大学肿瘤研究所,湖南长沙410078

出　　处：《医学临床研究》2007年第11期1855-1857,共3页Journal of Clinical Research

摘　　要：【目的】建立稳定分泌抗人心肌肌钙蛋白T(cardic troponin,cTnT)单克隆抗体的杂交瘤细胞株。【方法】以纯化的cTnT(分子质量为3.7×107)加福氏佐剂免疫8周龄雌性Balb/c小鼠,取免疫脾细胞与骨髓瘤细胞sp2/0融合,以酶联免疫吸附法(ELISA)筛选。阳性克隆以有限稀释法进行克隆化、Western blot法鉴定所制备的单克隆抗体。【结果】共筛出4株稳定分泌抗cTnT单克隆抗体的杂交瘤细胞株,所分泌的抗体效价高达10-6,经免疫学鉴定其分泌的免疫球蛋白类型均为IgG1、κ型,且Western blot鉴定4株细胞株所分泌的抗体均与人cTnT均有很好的反应性。【结论】获得4株稳定分泌抗人cTnT单克隆抗体的杂交瘤细胞株,为cTnT的临床研究提供了可能性。[Objective] To establish hybridoma cell lines that secrete anti-human troponin T （cTnT） monoclonal antibody（mAb） steadily. [Methods] Balb/c mice, female at eight-week age, were immunized with purified cTnT （MW 3. 7 × 10^7） and Freund adjuvant mixture by intraperitoneal. Spleen cells were fused with sp2/0 cells, and then selected by ELISA. Positive cells were cloned by limited dilution. Anti-human cTnT monoclonal antibodies（mAbs） secreted by the hybridoma cell lines were further confirmed by western blot. [Results] Four hybridoma cell lines secreting anti-human cTnT monoclonal antibody were established after cell fusion and limited dilution. The titer of antibodies secreted by four cell lines reached 10^-6 , and all antibodies were IgG1 subclass and κ type by immune identification. Western blot results showed that these four monoclonal antibodies reacted well with human cTnT. [Conclusion] Four hybridoma cell lines secreting mAbs against cTnT were produced, which provide the possibility for the clinical research of cTnT.

关 键 词：肌钙蛋白 杂交瘤 

分 类 号：R392.11[医药卫生—免疫学]