Spectroscopic and quartz crystal microbalance (QCM) characterisation of protein-based MIPs

El-Sharif, H.F., Aizawa, H. and Reddy, Subrayal M orcid iconORCID: 0000-0002-7362-184X (2015) Spectroscopic and quartz crystal microbalance (QCM) characterisation of protein-based MIPs. Sensors and Actuators, B: Chemical, 206 . pp. 239-245. ISSN 0925-4005

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Official URL: http://dx.doi.org/10.1016/j.snb.2014.09.053


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.

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