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首页> 外文期刊>Journal of Neuroimmunology: Official Bulletin of the Research Committee on Neuroimmunology of the World Federation of Neurology >Development of an ultrasensitive enzyme immunoassay for the determination of matrix metalloproteinase-9 (MMP-9) levels in normal human cerebrospinal fluid.
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Development of an ultrasensitive enzyme immunoassay for the determination of matrix metalloproteinase-9 (MMP-9) levels in normal human cerebrospinal fluid.

机译：用于确定正常人脑脊髓液中基质金属蛋白酶9（MMP-9）水平的超灵敏酶免疫测定法的开发。

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Determination of matrix metalloproteinase-9 (MMP-9) in human cerebrospinal fluid (CSF) to study blood-brain barrier impairment and immune cell migration in inflammatory neurological diseases recently became a matter of major interest. Regularly, MMP-9 was determined qualitatively or semi-quantitatively by zymography (gelatin gel electrophoresis) or quantitatively by enzyme immunoassay (EIA). As yet, it was not possible by either method to detect MMP-9 in CSF of controls (patients without pathologically increased CSF parameters). We developed an ultrasensitive two-side enzyme-linked immunosorbent assay (ELISA) which allows for the first time to measure reliably MMP-9 concentrations in CSF of controls. This ELISA uses a monoclonal as capture and a polyclonal as detector antibody. The detection limit of the assay is below 10 pg/ml and the assay range is 15-2000 pg/ml. Intra-assay precision is 2.5% for low and 3.7% for high, inter-assay precision is 11% for low and 10.7% for high values, respectively. The determination of the MMP-9 concentration in 50 control CSF gave the following results: range, 22-146 pg/ml; median, 76 pg/ml. The measurement of native and recombinant MMP-9 was carried out with three commercially available ELISAs, most widely employed in MMP-9 research, and compared to the newly developed one. All ELISAs recognize recombinant MMP-9 by factors of 5-20 less sensitively than native MMP-9.
机译：确定人类脑脊液（CSF）中的基质金属蛋白酶9（MMP-9）以研究炎性神经疾病中的血脑屏障损害和免疫细胞迁移成为最近引起人们关注的问题。通常，通过酶谱法（明胶凝胶电泳）或通过酶免疫法（EIA）定量或半定量测定MMP-9。迄今为止，通过这两种方法都无法检测到对照（没有病理性脑脊液参数升高的患者）的脑脊液中MMP-9。我们开发了一种超灵敏的两侧酶联免疫吸附试验（ELISA），该方法首次可以可靠地测量对照CSF中MMP-9的浓度。此ELISA使用单克隆抗体作为捕获剂，使用多克隆抗体作为检测器抗体。该测定的检出限低于10 pg / ml，测定范围为15-2000 pg / ml。批内精确度为低值的2.5％和高3.7％，批间精密度为低值的11％和高值的10.7％。测定50个对照脑脊液中MMP-9的浓度可得出以下结果：范围22-146 pg / ml;中位数为76 pg / ml。天然和重组MMP-9的测量是使用三种市售ELISA（最广泛用于MMP-9研究）进行的，与新开发的ELISA比较。所有ELISA识别重组MMP-9的灵敏度都比天然MMP-9低5-20倍。

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