State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, PR China.
Anal Chim Acta. 2012 Apr 20;723:45-53. doi: 10.1016/j.aca.2012.02.020. Epub 2012 Feb 19.
A novel temperature-sensitive molecular imprinted hydrogel composed of 2-acrylamido-2-methyl-propanosulfonic acid (AMPS), N-isopropylacrylamide (NIPAm) and acrylamide (AAm) has been prepared by free-radical cross-linking copolymerization in aqueous solution under two different temperatures (25 °C and -20 °C). Bovine serum albumin (BSA, pI 4.9, MW 66.0 kDa) is used as the template protein. The influence of the external temperature stimuli on the affinity of the hydrogels was investigated, and the optimal binding conditions were tested. The adsorption capacity (Q(max)) and association constant (K) for the specific interaction between the hydrogel and the template protein were determined by Langmuir isotherm plots. Several types of reference protein, which are different in molecular weights and isoelectric points were chosen to investigate the selectivity of the hydrogels. It was shown that the shape memory and the charge effect were the major factors for the recognition. This imprinted hydrogel was used to specifically adsorb the BSA from the protein mixture and real sample, which demonstrated its potential selectivity.
一种新型的温敏分子印迹水凝胶由 2-丙烯酰胺-2-甲基丙磺酸（AMPS）、N-异丙基丙烯酰胺（NIPAm）和丙烯酰胺（AAm）组成，通过在两种不同温度（25°C 和-20°C）下的水溶液自由基交联共聚制备。牛血清白蛋白（BSA，pI 4.9，MW 66.0 kDa）用作模板蛋白。研究了外部温度刺激对水凝胶亲和力的影响，并测试了最佳结合条件。通过 Langmuir 等温线图确定了水凝胶与模板蛋白之间特异性相互作用的吸附容量（Q(max)）和结合常数（K）。选择了几种分子量和等电点不同的参考蛋白来研究水凝胶的选择性。结果表明，形状记忆和电荷效应是识别的主要因素。该印迹水凝胶可用于从蛋白质混合物和实际样品中特异性吸附 BSA，证明了其潜在的选择性。
