The purification of cardiac troponin T(cTnT) from human left vertricular muscle has been developed. The 5 mg cTnT were obtained from 100 g cardiac muscle by homogenization, 70℃ treatment, imidazol-HCl dialysis, and DEAE-cellulose chromatography. The purity of total cTnT was 97.6 %. The cTnT was used to immunize Balb/C mice by intraspleenic injection and five hybridoma cell lines (G₃, G₈, G₁₀, A₅, A₇), which secreted stably monoclonal antibody against human cTnT, were obtained by cell fusion, identification, cloning, four lines were IgM, one line IgG by immunological analysis. The number of hybridomas chromosomes were 92～110, the McAb titers from ascitic fluid were about 3.2×10^-6～1. 6×10^-7.