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Immobilized Enzyme Reactor Chromatography for Online Gelatinase Inhibitors Screening

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Abstract

In this work, a new immobilized enzyme reactor (IMER) containing gelatinase enzyme (a mixture of MMP-2 and -9) was developed and then inserted into a HPLC system to evaluate the inhibitory potency of novel compounds towards gelatinase. The gelatinase was immobilized on an epoxy-silica column by in situ technique. To monitor the performance of the newly developed method, the kinetic constant (K m) of immobilized gelatinase as well as the inhibition constant (IC50) of a known inhibitor, i.e., LY52, were determined and these results were compared with those obtained by classical succinyl-gelatin assay. The IMER-HPLC system was subsequently applied to the screening of 39 synthetic compounds. The inhibitory potential of these inhibitors (expressed in IC50) was found to be in agreement with the literature.

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Acknowledgments

This work was financially supported by the Shandong Provincial Natural Science Foundation of China (2009ZRB02230).

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The authors have declared no conflict of interest.

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Authors and Affiliations

  1. School of Pharmaceutical Sciences, Shandong University, Jinan, 250012, People’s Republic of China

    Weina Liang, Zhun Hou, Hairong Wang, Wenfang Xu & Weihong Wang

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  1. Weina Liang
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  2. Zhun Hou
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  3. Hairong Wang
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  4. Wenfang Xu
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  5. Weihong Wang
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Correspondence to Weihong Wang.

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Liang, W., Hou, Z., Wang, H. et al. Immobilized Enzyme Reactor Chromatography for Online Gelatinase Inhibitors Screening. Chromatographia 78, 763–773 (2015). https://doi.org/10.1007/s10337-015-2904-0

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  • DOI: https://doi.org/10.1007/s10337-015-2904-0

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