Aksaray University, Faculty of Arts and Science, Chemistry Department, Aksaray, Turkey.
J Chromatogr A. 2011 Jan 21;1218(3):484-90. doi: 10.1016/j.chroma.2010.11.074. Epub 2010 Dec 5.
The aim of this study is to prepare supermacroporous cryogels embedded with Cu(2+)-attached sporopollenin particles (Cu(2+)-ASP) having large surface area for high protein adsorption capacity. Supermacroporous poly(2-hydroxyethyl methacrylate) (PHEMA)-based monolithic cryogel column embedded with Cu(2+)-ASP was prepared by radical cryo-copolymerization of 2-hydroxyethyl methacrylate (HEMA) with N,N'-methylene-bis-acrylamide (MBAAm) as cross-linker directly in a plastic syringe for affinity purification of human serum albumin (HSA). Firstly, Cu(2+) ions were attached to sporopollenin particles (SP), then the supermacroporous PHEMA cryogel with embedded Cu(2+)-ASP was produced by free radical polymerization using N,N,N',N'-tetramethylene diamine (TEMED) and ammonium persulfate (APS) as initiator/activator pair in an ice bath. Embedded particles (10 mg) in PHEMA-based cryogel column were used in the adsorption/desorption of HSA from aqueous solutions. Optimum conditions of adsorption experiments were performed at pH 8.0 phosphate buffer, with flow rate of 0.5 mL/min, and at 5°C. The maximum amount of HSA adsorption from aqueous solution was very high (677.4 mg/g SP) with initial concentration 6 mg/mL. It was observed that HSA could be repeatedly adsorbed and desorbed to the embedded Cu(2+)-ASP in PHEMA cryogel without significant loss of adsorption capacity.
本研究旨在制备具有大表面积的超微孔冷冻凝胶，其中嵌入了具有高蛋白质吸附能力的 Cu(2+) 附着的孢粉素颗粒（Cu(2+)-ASP）。通过自由基冷冻共聚直接在塑料注射器中，将 2-羟乙基甲基丙烯酸酯（HEMA）与 N,N'-亚甲基双丙烯酰胺（MBAAm）作为交联剂聚合，制备了嵌入 Cu(2+)-ASP 的超微孔聚（2-羟乙基甲基丙烯酸酯）（PHEMA）基整体式冷冻凝胶柱，用于亲和纯化人血清白蛋白（HSA）。首先，将 Cu(2+) 离子附着到孢粉素颗粒（SP）上，然后通过自由基聚合，在冰浴中使用 N,N,N',N'-四甲基乙二胺（TEMED）和过硫酸铵（APS）作为引发剂/活化剂对 SP 进行聚合，生成嵌入 Cu(2+)-ASP 的超微孔 PHEMA 冷冻凝胶。将嵌入的颗粒（10mg）装入 PHEMA 基冷冻凝胶柱中，用于从水溶液中吸附/解吸 HSA。在 pH8.0 的磷酸盐缓冲液中，流速为 0.5mL/min，温度为 5°C 时，吸附实验的最佳条件为吸附。从初始浓度为 6mg/mL 的水溶液中，HSA 的最大吸附量非常高（677.4mg/g SP）。观察到 HSA 可以反复吸附和解吸到嵌入 PHEMA 冷冻凝胶中的 Cu(2+)-ASP，而吸附容量没有明显损失。
