基质金属蛋白酶定量凝胶酶谱法的重新评估
Reappraisal of quantitative gel zymography for matrix metalloproteinases.
作者信息
Prescimone Tommaso, Tognotti Danika, Caselli Chiara, Cabiati Manuela, D'Amico Andrea, Del Ry Silvia, Giannessi Daniela
机构信息
Consiglio Nazionale delle Ricerche (CNR), Institute of Clinical Physiology, Laboratory of Cardiovascular Biochemistry, Pisa, Italy.
出版信息
J Clin Lab Anal. 2014 Sep;28(5):374-80. doi: 10.1002/jcla.21696. Epub 2014 Mar 19.
BACKGROUND
The determination of matrix metalloproteases (MMPs) is relevant in many pathophysiological conditions, especially if associated with extracellular matrix remodeling; however, the results obtained are closely linked to the method used and are not directly comparable. The aim of this study was to perform a reappraisal of quantitative gel zymography technique for MMPs in human plasma, to use for comparison with commercially available ELISA and in those experimental conditions where the MMP active form needs to be revealed.
METHODS
The critical methodological parameters of zymography were checked and a comparison with a routinely used ELISA was performed.
RESULTS
Sensitivity and reproducibility levels of zymography are suitable for detection of MMP-9 in human plasma, providing results closely related to those obtained by ELISA.
CONCLUSIONS
Analytical parameters of zymography were suitable for detection of MMPs in human plasma. Quantitative zymography for MMPs is an alternative method for comparing the results of ELISA widely employed for MMP determination, thus reducing the discrepancies between laboratories regarding gelatinase assay.
背景
基质金属蛋白酶（MMPs）的测定在许多病理生理状况中具有重要意义，特别是当与细胞外基质重塑相关时；然而，所获得的结果与所使用的方法密切相关，且无法直接进行比较。本研究的目的是对用于检测人血浆中MMPs的定量凝胶酶谱技术进行重新评估，以便与市售酶联免疫吸附测定（ELISA）进行比较，并用于那些需要揭示MMP活性形式的实验条件。
方法
检查了酶谱法的关键方法学参数，并与常规使用的ELISA进行了比较。
结果
酶谱法的灵敏度和重现性水平适用于检测人血浆中的MMP-9，所提供的结果与ELISA法获得的结果密切相关。
结论
酶谱法的分析参数适用于检测人血浆中的MMPs。MMPs的定量酶谱法是一种替代方法，可用于比较广泛用于MMP测定的ELISA结果，从而减少实验室之间在明胶酶测定方面的差异。
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