Department of Chemistry, Faculty of Engineering and Physical Sciences, University of Surrey, GU2 7XH, United Kingdom.
Biomacromolecules. 2012 Dec 10;13(12):3959-65. doi: 10.1021/bm301189f. Epub 2012 Nov 9.
We have characterized the imprinting capability of a family of acrylamide polymer-based molecularly imprinted polymers (MIPs) for bovine hemoglobin (BHb) and trypsin (Tryp) using spectrophotometric and quartz crystal microbalance (QCM) sensor techniques. Bulk gel characterization on acrylamide (AA), N-hydroxymethylacrylamide (NHMA), and N-isopropylacrylamide (NiPAM) gave varied selectivities when compared with nonimprinted polymers. We have also harnessed the ability of the MIPs to facilitate protein crystallization as a means of evaluating their selectivity for cognate and noncognate proteins. Crystallization trials indicated improved crystal formation in the order NiPAM<AA<NHMA. QCM studies of thin film MIPs confirm this trend with N-hydroxymethyl acrylamide MIPs exhibiting best discrimination between MIP and NIP and also cognate/noncognate protein loading. Equivalent results for acrylamide MIPs suggested that the cavities were equally selective for both proteins, while N-isopropylacrylamide MIPs were not selective for either cognate BHb or noncognate BSA. All BHb MIP-QCM sensors based on AA, NHMA, or NiPAM were essentially nonresponsive to smaller, noncognate proteins. Protein crystallization studies validated the hydrophilic efficacy of MIPS indicated in the QCM studies.
我们使用分光光度法和石英晶体微天平（QCM）传感器技术，对一系列丙烯酰胺基分子印迹聚合物（MIPs）对牛血红蛋白（BHb）和胰蛋白酶（Tryp）的印迹能力进行了表征。与非印迹聚合物相比，丙烯酰胺（AA）、N-羟甲基丙烯酰胺（NHMA）和 N-异丙基丙烯酰胺（NiPAM）的整体凝胶特性具有不同的选择性。我们还利用 MIP 促进蛋白质结晶的能力，作为评估其对同源和非同源蛋白质选择性的一种手段。结晶试验表明，NiPAM<AA<NHMA 的结晶形成顺序得到改善。薄膜 MIP 的 QCM 研究证实了这一趋势，N-羟甲基丙烯酰胺 MIP 对 MIP 和 NIP 以及同源/非同源蛋白质负载表现出最佳的区分能力。丙烯酰胺 MIP 的等效结果表明，这些腔对两种蛋白质具有同等的选择性，而 N-异丙基丙烯酰胺 MIP 对同源的 BHb 或非同源的 BSA 均没有选择性。所有基于 AA、NHMA 或 NiPAM 的 BHb MIP-QCM 传感器对较小的、非同源蛋白质基本上没有反应。蛋白质结晶研究验证了 QCM 研究中表明的 MIPS 的亲水性效果。
