两性聚丙烯酰胺冰胶对蛋白质分子印迹的广泛适用性

选择溶菌酶、胃蛋白酶、卵清蛋白、牛血红蛋白和γ-球蛋白为模板,考察两性聚丙烯酰胺冰胶的分子印迹功能。预聚溶液中除了丙烯酰胺、甲叉双丙烯酰胺外,还含有作为功能单体的丙烯酸和烯丙基胺,因此所得聚合物中既有酸性基团,也有碱性基团,有助于在分子水平上识别同样是两性物质的蛋白质。上述蛋白质的分子量、等电点差异显著,且涵盖了很宽的范围。利用同一单体溶液即可合成对所有蛋白质均有印迹效果的聚合物,其分子量和等电点无影响。模板分子在毛细管印迹柱中的保留时间大于非印迹柱中的保留时间。溶菌酶的印迹因子高达7.0,γ-球蛋白最小(2.0),其它蛋白质介于二者之间。本研究结果表明,两性聚丙烯酰胺冰胶是印迹各种蛋白质的理想材料,在蛋白质的识别、纯化和去除方面具有极大的潜力。

Extensive Imprinting Adaptability of Polyacrylamide-based Amphoteric Cryogels Against Protein Molecules

Lysozyme, pepsin, ovalbumin, hemoglobin andγ-globumin were chosen as templates to investigate the imprinting capability of amphoteric polyacrylamide cryogels. Prepolymerizing solutions contained acrylic acid and allyl amine, as well as acrylamide and N,N'-methylenebisacrylamide as the functional monomers. As a result there were both acidic and basic functional groups in the polymers, facilitating effective interactions with likewise amphoteric proteins. The proteins differ greatly and cover wide scopes of molecular weights and isoelectric points. Regardless of the values of the molecular weights and isoelectric points, all the templates gave higher retentions on the MIP tubes than on the NIP tube. The MIP of lysozyme indicated the highest imprinting factor of 7. 0, and that of γ-globumin showed the lowest, 2. 0. The values of other proteins were intervenient. Conclusively the amphoteric polyacrylamide cryogels were suitable imprinting materials for various proteins, and could potentially be used for protein recognition, purification, and depletion.