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Reference type: Journal
Authors: El-Sharif HF, Aizawa H, Reddy SM
Article Title: Spectroscopic and quartz crystal microbalance (QCM) characterisation of protein-based MIPs.
Publication date: 2015
Journal: Sensors and Actuators B: Chemical
Volume: 206
Page numbers: 239-245.
DOI: 10.1016/j.snb.2014.09.053
Alternative URL: http://www.sciencedirect.com/science/article/pii/S0925400514011228

Abstract: We have studied acrylamide-based polymers of varying hydrophobicity (acrylamide, AA; N-hydroxymethylacrylamide, NHMA; N-isopropylacrylamide, NiPAm) for their capability of imprinting protein. Rebinding capacities (Q) from spectroscopic studies were highest for bovine haemoglobin (BHb) MIPs based on AA, Q = 4.8 0.21 < NHMA, Q = 4.3 0.32 < NiPAm, Q = 3.6 0.45, while also demonstrating low selectivities for non-template proteins (<30 5%), with the exception of bovine serum albumin (BSA, >76 0.5%). When applied to the QCM sensor as thin-film MIPs, NHMA MIPs were found to exhibit best discrimination between MIP and non-imprinted control polymer (NIP) in the order of NiPAm < AA < NHMA. The extent of template removal and rebinding, using both crystal impedance and frequency measurements, demonstrated that 10% (w/v):10% (v/v) sodium dodecyl sulphate:acetic acid (pH 2.8) was efficient at eluting template BHb (with 80 10% removal). Selectivity studies of NHMA BHb-MIPs revealed higher adsorption and selective recognition properties to BHb (64.5 kDa) when compared to non-cognate BSA (66 kDa), myoglobin (Mb, 17.5 kDa), lysozyme (Lyz, 14.7 kDa) thaumatin (Thau, 22 kDa) and trypsin (Tryp, 22.3 kDa). The QCM gave frequency shifts of ~1500 50 Hz for template BHb rebinding in both AA and NHMA MIPs, whereas AA-based MIPs exhibited an interference signal of ~2200 50 Hz for non-cognate BSA in comparison to a ~500 50 Hz shift with NHMA MIPs. Our results show that NHMA-based hydrogel MIP are superior to AA and NIPAM.
Template and target information: protein, bovine haemoglobin, BHb
Author keywords: molecular imprinted polymer (MIP), hydrogel, protein, biosensor, QCM


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