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.PDF

240 BULL HUNAN MED UNIV 2003, 28( 3) 1 侯 珏, 胡国龄, 刘双虎, 谭德明, 刘映霞, 沙新平 ( 中南大学湘雅医院传染病研究所, 长沙410078) [] 目的: 获得兔抗人基质金属蛋白酶1(MMP1) 多 隆抗体, 并将其初步应用于临床检测方法: 将人 MMP1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MMP1 多 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MMP1的OD 值结果 : 获得初步纯化的兔抗人MMP1特异性多 隆抗体, 该抗体能与人MMP1起反应结论 : 制备的MMP1 多 隆抗体可初步用于临床检测 [] 人基质金属蛋白酶1融合蛋白; 亲和层析纯化; 多 隆抗体 [] R575 [] A [ ] 2003 Purification of human MMP1fusion protein and clinical detection of MMP1 by its polyclonal antibody HOU Jue, HU uoling, LIU Shuanghu, et al . ( I nstitute of I nf ectious Diseases , X i angy a H osp ital , Central South Univers ity , Changsha 410008, China) Abstract: Objectives To obtain the fusion protein of MMP1and therabbitantihuman matrix metal loproteinase1(MMP1) polyclonal antibody, and to detect the serum MMP1in patients by this polyclonal an tibody. 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 (Ig ) obtained from the immune serawere purified by ammonium sulfate fractionation, and the indirect sandwich enzyme im munoassaywas used to detect the serum MMP1in 24 samples of patientswith chronicliver disease or cirrhosis and in 12 normal sera as controls. Results The purified MMP1 fusion protein was successfully obtained; the purified specific polyclonal antibodies of rabbitantihuman MMP1were also obtained from the immune sera of the rabbits, and could respond tohuman MMP1. Comparedwith