Spectroscopic and quartz crystal microbalance (QCM) characterisation of protein-based MIPs
Spectroscopic and quartz crystal microbalance (QCM) characterisation of protein-based MIPs
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DOI：
10.1016/j.snb.2014.09.053
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发表时间：
2015-01-01
影响因子：
8.4
通讯作者：
Reddy, Subrayal M.
中科院分区：
文献类型：
--
作者：
EL-Sharif, Hazim F.;Aizawa, Hidenbou;Reddy, Subrayal M.
我们研究了不同疏水性的丙烯酰胺基聚合物（丙烯酰胺，AA; N-羟甲基丙烯酰胺，NHMA; N-异丙基丙烯酰胺，NiPAm）的印迹蛋白质的能力。来自光谱研究的再结合能力（Q）对于基于AA的牛血红蛋白（BHb）MIP是最高的，Q = 4.8 +/-0.21 < NHMA，Q = 4.3 +/-0.32 < NiPAm，Q = 3.6 +/-0.45，同时还证明了对非模板蛋白的低选择性（76 +/-0.5%）。当作为薄膜分子印迹聚合物应用于QCM传感器时，发现NHMA分子印迹聚合物在MIP和非印迹对照聚合物（NIP）之间表现出最好的区分，顺序为NiPAm < AA < NHMA。使用晶体阻抗和频率测量的模板去除和再结合的程度证明，10%（w/v）：10%（v/v）十二烷基硫酸钠：乙酸（pH 2.8）在洗脱模板BHb（去除80 +/-10%）时是有效的。NHMA BHb-MIPs的选择性研究显示，与非同源BSA（66 kDa）、肌红蛋白（Mb，17.5 kDa）、溶菌酶（Lyz，14.7 kDa）、索马甜蛋白（Thau，22 kDa）和胰蛋白酶（Tryp，22.3 kDa）相比，对BHb（64.5 kDa）具有更高的吸附和选择性识别特性。QCM在AA和NHMA MIP中对于模板BHb再结合给出了类似于1500 +/-50 Hz的频移，而与NHMA MIP的类似于500 +/-50 Hz的频移相比，基于AA的MIP对于非同源BSA表现出类似于2200 +/-50 Hz的干扰信号。我们的研究结果表明，基于NHMA的水凝胶MIP是上级优于AA和NIPAM。(C)2014爱思唯尔有限公司版权所有。
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 (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 similar to 1500 +/- 50 Hz for template BHb rebinding in both AA and NHMA MIPs, whereas AA-based MIPs exhibited an interference signal of similar to 2200 +/- 50 Hz for non-cognate BSA in comparison to a similar to 500 +/- 50 Hz shift with NHMA MIPs. Our results show that NHMA-based hydrogel MIP are superior to AA and NIPAM. (C) 2014 Elsevier B.V. All rights reserved.