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Autor(en) / Beteiligte
Titel
Low-volume multiplexed proteolytic activity assay and inhibitor analysis through a pico-injector arrayElectronic supplementary information (ESI) available. See DOI: 10.1039/c4lc01162g
Erscheinungsjahr
2015-02
Link zum Volltext
Quelle
Alma/SFX Local Collection
Beschreibungen/Notizen
  • Secreted active proteases, from families of enzymes such as matrix metalloproteinases (MMPs) and ADAMs (a disintegrin and metalloproteinases), participate in diverse pathological processes. To simultaneously measure multiple specific protease activities, a series of parallel enzyme reactions combined with a series of inhibitor analyses for proteolytic activity matrix analysis (PrAMA) are essential but limited due to the sample quantity requirements and the complexity of performing multiple reactions. To address these issues, we developed a pico-injector array to generate 72 different reactions in picoliter-volume droplets by controlling the sequence of combinational injections, which allowed simultaneous recording of a wide range of multiple enzyme reactions and measurement of inhibitor effects using small sample volumes (~10 μL). Multiple MMP activities were simultaneously determined by 9 different substrates and 2 inhibitors using injections from a pico-injector array. Due to the advantages of inhibitor analysis, the MMP/ADAM activities of MDA-MB-231, a breast cancer cell line, were characterized with high MMP-2, MMP-3 and ADAM-10 activity. This platform could be customized for a wide range of applications that also require multiple reactions with inhibitor analysis to enhance the sensitivity by encapsulating different chemical sensors. In this study we developed a pico-injector array to generate 9 × 2 × 2 × 2 = 72 different reactions in picoliter-sized droplets by controlling the sequence of combinational injections, which allows simultaneous read-outs of a wide range of multiple protease reactions and measurement of inhibitor effects by using small sample volumes.
Sprache
Englisch
Identifikatoren
ISSN: 1473-0197
eISSN: 1473-0189
DOI: 10.1039/c4lc01162g
Titel-ID: cdi_rsc_primary_c4lc01162g
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