a Vocational School of Health Services, Department of Medical Laboratory Techniques , Izmir University of Economics , Izmir , Turkey.
b Faculty of Science, Department of Biochemistry , Ege University , Izmir , Turkey.
J Biomater Sci Polym Ed. 2018 Dec;29(18):2218-2236. doi: 10.1080/09205063.2018.1534423. Epub 2018 Nov 5.
In this presented study, a novel molecularly imprinted polymeric hydrogel membranes (PHMs) were developed to use for the albumin depletion studies. For this, albumin imprinted poly(2-hydroxyethyl methacrylate-N-methacryloyl-(L)-phenylalanine methyl ester) polymeric hydrogel membranes [p(HEMA-MAP) PHMs] were synthesized by the photopolymerization technique, and then characterized by SEM, EDX, FT-IR and swelling studies. Synthesized PHMs had spherical structure and the MAP monomer incorporation onto the PHMs was determined by EDX analysis by using nitrogen stoichiometry. Also, the swelling ratio of the albumin imprinted p(HEMA-MAP) PHMs was determined as 215%. The optimum albumin adsorption condition (adsorption capacity, medium pH, adsorption rate, temperature, ionic strength) were studied and the maximum albumin adsorption capacity was found to be as 34.28 mg/g PHMs. Selectivity experiments were also carried out with the presence of the competitive proteins such as lysozyme and amylase, and the results demonstrated that the albumin imprinted p(HEMA-MAP) PHMs showed high affinity towards the BSA molecules than the competitive proteins of lysozyme and amylase. Adsorbed albumin was desorbed from the PHMs by 1.0 M of NaCl, and the reusability of the imprinted PHMs was also demonstrated for five successive adsorption-desorption cycles without any significant loss in the albumin adsorption capacity. As an application, sodium-dodecyl sulfate polyacrylamide gel electrophoresis was used to indicate the albumin depletion efficiency of albumin imprinted p(HEMA-MAP) PHMs. This presented study showed that, these imprinted membranes are promising for proteomic studies and applications, and can be used for the investigations for human diagnostics.
在本研究中，开发了一种新型的分子印迹聚合物水凝胶膜（PHM），用于白蛋白耗尽研究。为此，通过光聚合技术合成了白蛋白印迹聚（2-羟乙基甲基丙烯酸酯-N-甲基丙烯酰基-（L）-苯丙氨酸甲酯）聚合物水凝胶膜[p（HEMA-MAP）PHM]，然后通过 SEM、EDX、FT-IR 和溶胀研究进行了表征。合成的 PHM 具有球形结构，通过 EDX 分析（使用氮化学计量法）确定 MAP 单体在 PHM 上的掺入。此外，白蛋白印迹 p（HEMA-MAP）PHM 的溶胀比确定为 215%。研究了白蛋白印迹 p（HEMA-MAP）PHM 的最佳白蛋白吸附条件（吸附容量、介质 pH、吸附速率、温度、离子强度），发现最大白蛋白吸附容量为 34.28mg/g PHM。还进行了选择性实验，存在竞争蛋白如溶菌酶和淀粉酶，结果表明，白蛋白印迹 p（HEMA-MAP）PHM 对 BSA 分子表现出比溶菌酶和淀粉酶等竞争蛋白更高的亲和力。通过 1.0 M 的 NaCl 从 PHM 上解吸吸附的白蛋白，并且还证明了印迹 PHM 的可重复使用性，在五个连续的吸附-解吸循环中没有白蛋白吸附容量的任何显著损失。作为应用，十二烷基硫酸钠聚丙烯酰胺凝胶电泳用于指示白蛋白印迹 p（HEMA-MAP）PHM 的白蛋白耗尽效率。本研究表明，这些印迹膜在蛋白质组学研究和应用中具有广阔的应用前景，可用于人类诊断的研究。
