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Covalent Binding of Antibodies to Cellulose Paper Discs and Their Applications in Naked-eye Colorimetric Immunoassays
Published on: October 21, 2016
Molecularly imprinted polymer modified g-C3N4 integrated into paper-based devices for colorimetric detection of
Phan Quang Huy Quang1,2,3,4, Huynh Anh Khoa2,5,3, Hoang Cao Nguyen2,5,3
1Faculty of Biology and Environment, Ho Chi Minh City University of Industry and Trade, 140 Le Trong Tan Street, Tay Thanh Ward, Ho Chi Minh City, Vietnam.
Abstract:
To enable rapid, on-site monitoring of amoxicillin and ensure controlled administration, we have developed a paper-based analytical device (PAD) incorporating a molecularly imprinted polymer (MIP) anchored on graphitic carbon nitride (g-C₃N₄) nanosheets. The MIP@g-C₃N₄ composite was synthesized via in situ polymerization of 3-aminopropyl triethoxysilane (APTES) and tetraethyl orthosilicate (TEOS) around amoxicillin templates on g-C₃N₄, yielding highly specific binding cavities upon template removal. The g-C₃N₄ nanozyme's intrinsic peroxidase-like activity catalyzes the oxidation of colorless 3,3',5,5'-tetramethylbenzidine (TMB) to its blue oxidized form (oxTMB) in the presence of hydrogen peroxide. In the absence of amoxicillin, a vivid blue signal develops, whereas competitive binding of amoxicillin at the MIP sites inhibits nanozyme activity, causing a proportional decrease in color intensity. Analytical characterization demonstrated a linear response over 0-100 µM amoxicillin, with a limit of detection (LOD) of 0.97 µM. The integration with the PAD format permits semi-quantitative visual readout and quantitative determination via a portable smartphone, all within minutes and without the need for complex instrumentation. When applied to spiked milk and tap water, the MIP@g-C₃N₄ sensor achieved recoveries of 92-105% with negligible interference from structurally related antibiotics. The marriage of MIP selectivity, g-C₃N₄ nanozyme catalysis, and low-cost PAD architecture delivers a user-friendly, field-deployable platform for rapid amoxicillin screening, promising significant potential to bolster antibiotic stewardship and mitigate overdose-related health risks across clinical, environmental, and food-safety applications.
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