Separation/enrichment of active natural low content protein using protein imprinted polymer

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• 作者：Ruifang Han ; Xiaocui Xing ; Ying Wang ; Yi Long ; Yang Sun ; Zhuo Zhao ; Huaifeng Mi
• 关键词：Protein-imprinted polymer ; Assistant recognition polymer chains (ARPCs) ; Separation of natural protein ; Cyclophilin 18 ; PPIase activity
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2008
• 出版时间：15 September 2008
• 年：2008
• 卷：873
• 期：1
• 页码：113-118
• 全文大小：499 K

文摘

We describe a new type of protein-imprinted polymer for separation/enrichment of active natural protein present at a relatively low level in cell extracts, with a cloned bacterial protein as template. In this work, cloned pig cyclophilin 18 (pCyP18) was used as template. The template protein was selectively assembled with assistant recognition polymer chains (ARPCs) from their library, which consists of numerous limited length polymer chains with randomly distributed recognition and immobilizing sites. These assemblies of protein and ARPCs were adsorbed by porous polymeric beads and immobilized by cross-linking polymerization. After removing the template, the synthesized imprinted polymer was used to adsorb authentic pCyP18 from cell extract, and its proportional content was enriched 200 times. The assay of peptidyl-prolyl cis–trans-isomerase (PPIase) activity showed that natural pCyP18 is more active than cloned pCyP18 and, in particular, it is much more sensitive to the suppressant cyclosporine A (CsA).