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https://hdl.handle.net/2440/94234
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Type: | Journal article |
Title: | Label-free real-time quantification of enzyme levels by interferometric spectroscopy combined with gelatin-modified nanoporous anodic alumina photonic films |
Author: | Nemati, M. Santos, A. Kumeria, T. Losic, D. |
Citation: | Analytical Chemistry, 2015; 87(17):9016-9024 |
Publisher: | American Chemical Society |
Issue Date: | 2015 |
ISSN: | 0003-2700 1520-6882 |
Statement of Responsibility: | Mahdieh Nemati, Abel Santos, Tushar Kumeria, and Dusan Losic |
Abstract: | Herein, 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. |
Keywords: | Aluminum Oxide Trypsin Gelatin Interferometry Electrodes Particle Size Surface Properties Photons Porosity Time Factors Nanostructures |
Rights: | © 2015 American Chemical Society |
DOI: | 10.1021/acs.analchem.5b02225 |
Grant ID: | http://purl.org/au-research/grants/arc/DE140100549 http://purl.org/au-research/grants/arc/DP120101680 http://purl.org/au-research/grants/arc/FT110100711 |
Published version: | http://dx.doi.org/10.1021/acs.analchem.5b02225 |
Appears in Collections: | Aurora harvest 2 Chemical Engineering publications |
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