富集纯化微量天然蛋白质的分子印迹聚合物.pdf

富集/纯化微量天然蛋白质的分子印迹聚合物* 宓怀风，赵琢，王春红，龙毅 （教育部功能高分子材料重点实验室，南开大学高分子化学研究所，天津300071 ） 摘要：利用分子印迹聚合技术，可以构造出对模板具有特定选择性识别位点的聚合物。除了小分子外，蛋白 质也可以作为印迹的模板。但是到目前为止，作为模板的蛋白质绝大部分是生命体中含量较高的蛋白质。在 这儿我们介绍一种新型的分子印迹聚合物合成方法，它用在细菌中克隆的蛋白质作为模板，用于富集/纯化细 胞提取液中微量的目标蛋白质。本工作中，克隆的亲环素18 （cyclophilin 18 ，CyP18 ）被用作模板。模板蛋白 质选择性地和辅助识别聚合物链进行组装。这些组装体被大孔吸附高分子微球吸附并被交联固定于其上。洗 脱蛋白质模板之后，合成的印迹聚合物用于吸附细胞提取液中的亲环素18，达1到了300倍的富集效果。 关键词：分子印迹聚合物；辅助识别聚合物链(ARPC)；克隆蛋白质作为模板；富集/纯化天然微量蛋白质 The molecular-imprinted polymer for enrichment/purification of non-abundant natural protein MI Huaifeng, ZHAO Zhuo, WANG Chunhong and LONG Yi Key Laboratory of Functional Polymer Materials, the Ministry of Education of China, Institute of Polymer Chemistry, Chemical School of Nankai UniversityTianjin, 300071, China Abstract: The molecular-imprinting technique creates specific recognition sites using template molecules. In addition to small molecules, proteins can also be used as templates. Up until now, only abundant proteins have been used as templates. Here we describe a new method for preparing a protein-imprinted polymer with a template of cloned bacterial protein for recognition/adsorption of the authentic target protein present at a relatively low level in cell extract. In this work, cloned pig cyclophilin 18 (pCyP18) was used as a template. This protein is ellipsoid in shape with molecular mass of 18 kDa. It makes up around 0.02–0.03% of the total cytosolic protein. The cloned pCyP18 was selectively assembled with assistant recognition polymer chains (ARPCs) from their library, which consists of numerous limited length polymer chains with randomly distributed recognition sites and immobilizing sites. The backbone of the ARPC, comprising limited length polyacrylic acids, was synthesized with a butyl acrylate