Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.jchromb.2010.04.040
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dc.titleOn-line chromatographic screening of matrix metalloproteinase inhibitors using immobilized MMP-9 enzyme reactor
dc.contributor.authorMa, X.
dc.contributor.authorChan, E.C.Y.
dc.date.accessioned2014-10-09T06:56:14Z
dc.date.available2014-10-09T06:56:14Z
dc.date.issued2010-07-01
dc.identifier.citationMa, X., Chan, E.C.Y. (2010-07-01). On-line chromatographic screening of matrix metalloproteinase inhibitors using immobilized MMP-9 enzyme reactor. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences 878 (21) : 1777-1783. ScholarBank@NUS Repository. https://doi.org/10.1016/j.jchromb.2010.04.040
dc.identifier.issn15700232
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/89651
dc.description.abstractMatrix metalloproteinase 9 (MMP-9) plays an important role in cancer invasion and metastasis and has been an attractive target for therapeutic intervention of cancer metastasis. However, considering the high cost and intricacy associated with the expression, isolation and purification of the recombinant enzyme for the screening of drug candidates, alternative methods that explore the recycling of enzymes become desirable. In this study, a new immobilized enzyme reactor (IMER) containing human recombinant MMP-9 enzyme was developed and characterized for the on-line screening of MMP-9 inhibitors. The MMP-9 IMER containing active unit of the enzyme (U = 0.08 μmol/min) on the disk was inserted into a HPLC system connected to a UV-vis detector for on-line chromatographic screening. The resulting conjugated enzyme was shown to be an active and stable catalyst for the hydrolysis of MMP-9 chromogenic thiopeptide substrate Ac-PLG-[2-mercapto-4-methyl-pentanoyl]-LG-OC2H5. The kinetics profile of the enzyme in IMER and free solution were determined and compared. Selected reversible MMP inhibitors, N-isobutyl-N-(4-methoxyphenylsulfonyl)-glycyl hydroxamic acid (NNGH), doxycycline and minocycline were further characterized using the MMP-9 IMER and free enzyme solution assays. Our system demonstrated applicability as a rapid and cost-effective screening tool for inhibitors of the MMP-9 enzyme. © 2010 Elsevier B.V. All rights reserved.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1016/j.jchromb.2010.04.040
dc.sourceScopus
dc.subjectHigh throughput screening
dc.subjectImmobilized enzyme reactor
dc.subjectMatrix metalloproteinase
dc.subjectMMP-9
dc.typeArticle
dc.contributor.departmentPHARMACY
dc.contributor.departmentCHEMICAL & BIOMOLECULAR ENGINEERING
dc.description.doi10.1016/j.jchromb.2010.04.040
dc.description.sourcetitleJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
dc.description.volume878
dc.description.issue21
dc.description.page1777-1783
dc.description.codenJCBAA
dc.identifier.isiut000279509300008
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