Label-free real-time quantification of enzyme levels by interferometric spectroscopy combined with gelatin-modified nanoporous anodic alumina photonic films

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dc.contributor.authorNemati, M.
dc.contributor.authorSantos, A.
dc.contributor.authorKumeria, T.
dc.contributor.authorLosic, D.
dc.date.issued2015
dc.description.abstractHerein, we present an interferometric sensor based on the combination of chemically functionalized nanoporous anodic alumina photonic films (NAA-PFs) and reflectometric interference spectroscopy (RIfS) aimed to detect trace levels of enzymes by selective digestion of gelatin. The fabrication and sensing performance of the proposed sensor were characterized in real-time by estimating the changes in effective optical thickness (i.e., sensing principle) of gelatin-modified NAA-PFs (i.e., sensing element) during enzymatic digestion. The working range (WR), sensitivity (S), low limit of detection (LLoD), and linearity (R(2)) of this enzymatic sensor were established by a series of experiments with different concentrations of gelatin (i.e., specific chemical sensing element) and trypsin (i.e., analyte), a model protease enzyme with relevant implications as a biomarker in the diagnosis of several diseases. The chemical selectivity of the sensor was demonstrated by comparison of gelatin digestion by other nonspecific enzyme models such as chymotrypsin and horseradish peroxidase. Furthermore, the role of the chemical sensing element (i.e., gelatin) was assessed by using hemoglobin instead of gelatin. Finally, we demonstrated that this sensor can be readily used to establish the kinetic parameters of enzymatic reactions. The obtained results revealed that the presented sensor has a promising potential to be used as a point-of-care system for fast detection of gastrointestinal diseases at early stages.
dc.description.statementofresponsibilityMahdieh Nemati, Abel Santos, Tushar Kumeria, and Dusan Losic
dc.identifier.citationAnalytical Chemistry, 2015; 87(17):9016-9024
dc.identifier.doi10.1021/acs.analchem.5b02225
dc.identifier.issn0003-2700
dc.identifier.issn1520-6882
dc.identifier.orcidSantos, A. [0000-0002-5081-5684]
dc.identifier.orcidLosic, D. [0000-0002-1930-072X]
dc.identifier.urihttp://hdl.handle.net/2440/94234
dc.language.isoen
dc.publisherAmerican Chemical Society
dc.relation.granthttp://purl.org/au-research/grants/arc/DE140100549
dc.relation.granthttp://purl.org/au-research/grants/arc/DP120101680
dc.relation.granthttp://purl.org/au-research/grants/arc/FT110100711
dc.rights© 2015 American Chemical Society
dc.source.urihttps://doi.org/10.1021/acs.analchem.5b02225
dc.subjectAluminum Oxide
dc.subjectTrypsin
dc.subjectGelatin
dc.subjectInterferometry
dc.subjectElectrodes
dc.subjectParticle Size
dc.subjectSurface Properties
dc.subjectPhotons
dc.subjectPorosity
dc.subjectTime Factors
dc.subjectNanostructures
dc.titleLabel-free real-time quantification of enzyme levels by interferometric spectroscopy combined with gelatin-modified nanoporous anodic alumina photonic films
dc.typeJournal article
pubs.publication-statusPublished

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