Studies on the synthesis and behavior of acrylate polymer stationary phases for cation exchange chromatography of proteins
Abstract
A new type of silica-based cation exchange packing material was synthesized and characterized using LiChrospher 1000 Diol (10$\mu$m, 1000A). The effect of acrylate monomer and Ce(iv) concentrations on the nature of the polymeric surface coating was investigated. The optimum amount of Ce(iv) required to initiate the maximum number of reaction sites was found to be 0.5 g/g of silica. The structure of this acrylate coating was investigated by detaching the coating from the silica surface and analyzing the acrylate polymer by mass spectrometry. MS data indicates that the coating is not a linear polymer. Ion exchange capacities of this packing material are 3-5 times higher than conventional type ion exchangers. Quantitation of ionic displacement revealed that the acrylate polymer does not wrap around the protein during the adsorption-desorption process. It was concluded that the enhanced capacities of these fimbriated coatings may be attributed to long distance electrostatic interactions. Proteins immobilized on these sorbents showed selectivity that was different than the ion exchange alone and was uniquely related to each protein. Lysozyme bearing columns showed entirely different selectivity toward some basic proteins. The ion exchange mechanism of these new packing material is not completely understood. The data indicates that these proteins immobilized columns have mixed mode effect.
Degree
Ph.D.
Advisors
Regnier, Purdue University.
Subject Area
Organic chemistry|Analytical chemistry
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