申请人:——
公开号:US05470916A1
公开(公告)日:1995-11-28
Matrices for electrokinetic and chromatographic separations, based on a unique class of mono- and disubstituted (on the amido nitrogen) acrylamides (e.g., acrylamido-N-ethoxy ethanol, acrylamido-N,N-diethoxy ethanol) off erthe following advantages: a) strong resistance to the alkaline hydrolysis (most zone separations occurring at basic pH values); b) a high hydrophilicity; and c) greater porosity, due to the larger M.sub.r value of the monomers. Such matrices can be prepared as `chemically crosslinked` gels or as `physical` gels, the latter consisting of strings of monofunctional monomer in the absence of crosslinker, at concentrations above the entanglement threshold. It is moreover possible to further increment the gel porosity by polymerizing it in the presence of laterally aggregating agents (e.g., polyethylene glycol of M.sub.r 6-20,000 Da), thus obtaining matrices which allow better resolution of large M.sub.r proteins and of nucleic acid fragments; by polymerizing the matrix in the presence of a gradient of the laterally aggregating agent, it is possible to obtain a porosity gradient even at a constant percent of monomers in the gel. Finally, photopolymerization (e.g., with methylene blue and a redox couple, sodium toluene sulfinate and diphenyliodonium chloride and light), instead with the standard red/ox catalysts, prevents formation of N-oxides of amines present in the polymerizing solution.
基于一类独特的单取代和双取代(在酰胺氮上)丙烯酰胺(例如,丙烯酰胺-N-乙氧乙醇,丙烯酰胺-N,N-二乙氧乙醇)的电动力学和色谱分离矩阵具有以下优点:a)强抗碱水解作用(大多数区分离发生在碱性pH值下);b)高亲水性;c)更大的孔隙度,由于单体的较大M.sub.r值。这样的矩阵可以制备为“化学交联”凝胶或“物理”凝胶,后者在超过纠缠阈值的浓度下由单官能单体串成,没有交联剂。此外,通过在侧向聚集剂(例如M.sub.r为6-20,000 Da的聚乙二醇)存在下聚合凝胶,可以进一步增加凝胶孔隙度,从而获得允许更好分离大M.sub.r蛋白质和核酸片段的矩阵;通过在侧向聚集剂的梯度存在下聚合基质,甚至可以在凝胶中单体的恒定百分比下获得孔隙度梯度。最后,与标准的氧化还原催化剂不同,光聚合(例如,用亚甲基蓝和氧化还原偶合物,甲苯磺酸钠和二苯基碘化铵和光)可防止聚合溶液中存在的胺的N-氧化物的形成。