以环糊精准聚轮烷为“假载体”印迹识别蛋白质的研究

Protein Imprinted with Cyclodextrin Pseudo-polyrotaxanes as Pseudo-supports

实验以聚乙烯醇分子为客体分子,与γ-环糊精(γ-CD)分子组装制备环糊精准聚轮烷(CD-PPRs);在金属离子存在下,以丙烯酰胺(AM)为单体,N,N-亚甲基双丙烯酰胺为交联剂,以CD-PPRs为"假载体",制备了以牛血清白蛋白(BSA)为模板的分子印迹聚合物(cp-MIP)。结果表明,随着功能基化CD-PPRs(CD-PPRsAc)用量增加,印迹聚合物吸附量先增加后减小,当CD-PPRs-Ac与AM的质量比为0.55时,印迹聚合物的吸附量最大;在Cu2+共存时,吸附量最高可达5.16 mg/g;将cp-MIP用于4种混合蛋白溶液吸附,结果表明,引入CD-PPRs使印迹聚合物的特异性吸附能力明显提高,特异性吸附量可提高约6倍。

Protein molecularly imprinted polymers （MIPs） have been prepared with cyclodextrin pseudo-polyrotaxanes （CD-PPRs） as pseudo-supports, acrylamide as the monomer and N,N-methylene bisacrylamide as the cross-linker. Firstly, CD-PPRs were obtained with T-cyclodextrin as hosts and poly （vinyl alcohol） as guests. Then CD-PPRs were modified with acryloyl chloride. Subsequently, the cp-MIP was prepared in the presence of the modified CD-PPRs （CD-PPRs-Ac） and metal ions with bovine serum albumin （BSA） as the template. The MIPs adsorption amount to the BSA template increased initially and then decreased with the increase content of CD-PPRs. It was found that the adsorption capacity of MIPs reached maximum when the ratio of CD-PPRs-Ac to AM was about 0.55. The adsorption maximum reached to 5.16 mg/g in the presence of certain amount of Cu2＋. Finally, the imprinted polymers were used to purify the template protein from the protein mixtures containing four （ BSA, ovalbumin, soybean trypsine inhibitor and lysozyme） different proteins to evaluate the adsorption specificity of MIPs. The experiments showed that the recognition specificity was improved obviously and the adsorption capacity increased by 6 times with the introduction of CD-PPRs.