胰蛋白酶分子表面印迹材料的制备及其大分子识别特性

Preparation of Trypsin Molecularly Surface-imprinted Material and Its Macromolecular Recognition Characteristics

以交联聚乙烯醇(CPVA)微球为基质,采用接枝聚合和表面印迹同步技术制备碱性蛋白胰蛋白酶(TRY)分子表面印迹材料,甲基丙烯酰氯与CPVA微球表面的羟基发生快速酯化反应,得到表面含大量可聚合双键甲基丙烯酰基(MAO)的改性微球MAO-CPVA.按一定摩尔比将TRY和单体阴离子单体对苯乙烯磺酸钠(SSS)溶解在水溶液中,加入交联剂N,N'-亚甲基双丙烯酰胺(MBA),MAO-CPVA分散于水介质中,过硫酸铵/亚硫酸氢钠引发体系产生自由基,使包围在TRY周围的单体SSS与MBA在MAO-CPVA表面发生接枝交联聚合,制得TRY表面印迹微球MIP-PSSS/CPVA,对其进行表征,考察了其大分子识别性能.结果表明,MIP-PSSS/CPVA对TRY有优良的亲和性和特异识别选择性,吸附容量达85.9 mg/g,对TRY的选择性系数相对于蛋白溶菌酶LZM达17.52.

Trypsin（TRY） molecularly surface-imprinted material was prepared by synchronous grafting polymerization and surface imprinting with crosslinked polyvinyl alcohol（CPVA） microspheres as matrix. The reaction between methyl acryloyl chloride and the hydroxyl groups on the surface of CPVA microspheres occurred quickly, resulting in the modified MAO-CPVA microspheres with a great deal of polymerizable double-bond methylacryloyl（MAO） group. Anionic monomer sodium p-styrene sulfonate（SSS） and template TRY in a certain molar ratio were added in an aqueous solution, interacting each other. And the crosslinker N, N＇-methylene bisacrylamide（MBA） was further added, and the modified MAO-CPVA microspheres were dispersed in the aqueous medium, the free radicals were produced by ammonium persulfate and sodium bisulfite redox initiating system, the graft-crosslinking polymerization of SSS surrounded around the template TRY and crosslinker MBA was conducted so that TRY surface-imprinted material MIP-PSSS/CPVA microspheres were obtained, and characterized. Moreover, their macromolecular recognition characteristics were studied. The results show that the MIP-PSSS/CPVA microspheres possess excellent binding affinity and specific recognition selectivity for the template TRY. The adsorption capacity of TRY on the microspheres reaches up to 85.9 mg/g, and the selectivity coefficient of imprinted TRY in contrast to protein lysozyme is up to 17.52.