Abstract
Capillary isoelectric focusing (CIEF) was first performed in an open tubular mode. It depended on an immobilized pH gradients from the polymerization of several solutions, which contained acrylamide, N,N'-methylenebisacrylamide, allylamine, and acrylic acid. Different ratios of the basic and acidic monomers lead to solutions of various pHs. When injected into a capillary in the sequence of their pHs, the solutions polymerized and resulted into polymers with a series of isoelectric points (pIs). These polymers formed a covalently attached hydrophilic layer in the capillary, acting as both column bonding and carrier ampholytes. When five polymers were orderly lined up in a capillary, an immobilized pH gradient was established. This open tubular column was subjected to CIEF, showing excellent performance properties for protein focusing.
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