Aslıyüce Sevgi, Mattiasson Bo, Denizli Adil
Department of Chemistry, Hacettepe University, Ankara, Turkey.
Department of Biotechnology, Lund University, Lund, Sweden.
Biomed Chromatogr. 2019 Oct;33(10):e4605. doi: 10.1002/bmc.4605. Epub 2019 Jul 3.
Cryogels have been demonstrated to be efficient when applied for protein isolation. Owing to their macroporous structure, cryogels can also be used for treating particle-containing material, e.g. cell homogenates. Another challenging development in protein purification technology is the use of molecularly imprinted polymers (MIPs). These MIPs are robust and can be used repeatedly. The paper presents a new technology that combine the formation of cryogel beads concomitantly with making imprints of a protein. Protein A was chosen as the print molecule which was also be the target in the purification step. The present paper describes a new method to produce protein-imprinted cryogel beads. The protein-imprinted material was characterized and the separation properties were evaluated with regard to both the target protein and whole cells with target protein exposed on the cell surface. The maximum protein A adsorption was 18.1 mg/g of wet cryogel beads. The selectivity coefficient of protein A-imprinted cryogel beads for protein A was 5.44 and 12.56 times greater than for the Fc fragment of IgG and protein G, respectively.
冷冻凝胶已被证明在用于蛋白质分离时是有效的。由于其大孔结构，冷冻凝胶还可用于处理含颗粒物质，如细胞匀浆。蛋白质纯化技术的另一个具有挑战性的进展是分子印迹聚合物（MIPs）的使用。这些MIPs性能稳定，可重复使用。本文介绍了一种新技术，该技术将冷冻凝胶珠的形成与蛋白质印迹同时进行。选择蛋白A作为印迹分子，其在纯化步骤中也是目标物。本文描述了一种生产蛋白质印迹冷冻凝胶珠的新方法。对蛋白质印迹材料进行了表征，并针对目标蛋白质以及细胞表面暴露有目标蛋白质的全细胞评估了分离性能。湿冷冻凝胶珠对蛋白A的最大吸附量为18.1 mg/g。蛋白A印迹冷冻凝胶珠对蛋白A的选择性系数分别比对IgG的Fc片段和蛋白G高5.44倍和12.56倍。
