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

Hunan yi ke da xue xue bao = Hunan yike daxue xuebao = Bulletin of Hunan Medical University Pub Date : 2003-06-01

Jue Hou, Guo-ling Hu, Shuang-hu Liu

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Abstract

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.

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[人MMP-1融合蛋白的纯化及其多克隆抗体的临床检测]。

目的:获得MMP-1与兔抗人基质金属蛋白酶-1(MMP-1)多克隆抗体的融合蛋白，并利用该多克隆抗体检测患者血清中MMP-1的含量。方法:采用亲和层析法纯化融合蛋白。用纯化的融合蛋白免疫2只家兔，采集家兔免疫血清。采用硫酸铵分离纯化免疫血清中抗体(IgG)，采用间接夹心酶免疫分析法检测24例慢性肝病或肝硬化患者和12例正常人血清中MMP-1的含量。结果:成功获得纯化的MMP-1融合蛋白;从兔免疫血清中获得纯化的兔抗人MMP-1特异性多克隆抗体，并能对人MMP-1产生应答。慢性肝炎、肝硬化患者血清MMP-1 OD值较正常对照组显著升高(P < 0.01)。结论:兔抗人MMP-1多克隆抗体可用于肝纤维化早期诊断。

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Hunan yi ke da xue xue bao = Hunan yike daxue xuebao = Bulletin of Hunan Medical University

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