人心肌肌钙蛋白T的纯化和单克隆抗体的制备

Purification of Human Cardiac Troponin T and Production Monoclonal Antibody Against It

从人左室心肌中成功纯化心肌肌钙蛋白Ｔ（ｃＴｎＴ）．经匀浆，７０℃加热处理，咪唑盐酸透析，ＤＥＡＥ－纤维素层析，１００ｇ心肌获取ｃＴｎＴ５ｍｇ，纯度为９７．６％．同时采用脾内免疫法，免疫Ｂａｌｂ／Ｃ小鼠，经细胞融合，筛选，克隆化得５株稳定分泌抗人ｃＴｎＴ单克隆抗体（ＭｃＡｂ）的杂交瘤细胞（Ｇ３，Ｇ８，Ｇ１０，Ａ５，Ａ７），４株为ＩｇＭ，１株为ＩｇＧ，染色体数目９２～１１０条．腹水效价为３．２×１０－６～１．６×１０－７．

The purification of cardiac troponin T(cTnT) from human left vertricular muscle hasbeen developed. The 5 mg cTnT were obtainedfrom 100 g cardiac muscle by homogenization,70t treatment, imidazol-HCl dialysis, andDEAE-cellulose chromatography. The purity oftotal cTnT was 97. 6 %. The cTnT was used toimmunize Balb/C mice by intraspleenic injectionand five hybridoma cell lines (G3, G8, G10 A5A7), which secreted stably monoclonal antibodyagainst human cTnT, were obtained by cellfusion, identification, cloning, four lines wereIgM, one line IgG by immunological analysis.The number of hybridomas chromosomes were92 - 110, the McAb titers from ascitic fluidwere about 3.2 × 10-6～ 1. 6×10-7.