人基质金属蛋白酶-1融合蛋白的纯化及其多克隆抗体的制备与初步应用

Purification of human MMP-1 fusion protein and clinical detection of MMP-1 by its polyclonal antibody

目的 :获得兔抗人基质金属蛋白酶 1 (MMP 1 )多克隆抗体 ,并将其初步应用于临床检测。方法 :将人MMP 1融合蛋白经亲和层析法纯化 ,并将其作为抗原免疫家兔 ,获得兔抗血清 ,经饱和硫酸铵纯化 ,获得初步纯化的兔抗人MMP 1多克隆抗体。用双抗夹心间接酶联免疫吸附实验 (ELISA)测定正常对照、慢性肝病、肝硬化患者血清MMP 1的OD值。结果 :获得初步纯化的兔抗人MMP 1特异性多克隆抗体 ,该抗体能与人MMP 1起反应。结论:制备的MMP

Objectives To obtain the fusion protein of MMP 1 and the rabbit anti human matrix metalloproteinase 1(MMP 1) polyclonal antibody, and to detect the serum MMP 1 in patients by this polyclonal antibody. Methods The fusion protein was purified by affinity chromatography.Two rabbits were immunized with the purified fusion protein, and the immune sera of rabbits were collected. Antibodies (IgG) obtained from the immune sera were purified by ammonium sulfate fractionation, and the indirect sandwich enzyme immunoassay was used to detect the serum MMP 1 in 24 samples of patients with chronic liver disease or cirrhosis and in 12 normal sera as controls. Results The purified MMP 1 fusion protein was successfully obtained; the purified specific polyclonal antibodies of rabbit anti human MMP 1 were also obtained from the immune sera of the rabbits, and could respond to human MMP 1. Compared with the normal controls, the OD value of serum MMP 1 significantly increased in patients with chronic hepatitis or liver cirrhosis ( P <0.01). Conclusion The rabbit anti human MMP 1 polyclonal antibody may be used in the diagnosis of initial stage of hepatic fibrosis.