Preparation of dye–ligand affinity chromatographic packings based on monodisperse poly(glycidylmethacrylate-co-ethylenedimethacrylate) beads and their chromatographic properties

Monodisperse porous particles, poly(glycidylmethacrylate-co-ethylenedimethacrylate), P(GMA-EDMA) beads with diameter of 7 μm were prepared by a single-step swelling and polymerization method. The polymer particles were advantaged through immobilization of Procion Bule MX-R, which was incubated by epichlorohydrin via the epoxide groups on the particles surface. The Procion Bule MX-R-immobilized P(GMA-EDMA) beads were mechanically stable and acted as the rigid matrix for column chromatography in HPLC mode. The chromatographic properties of the dye–ligand affinity chromatographic stationary phase for biopolymers separation are discussed. This affinity column has advantages of enabling biopolymer separation, high efficiency and low backpressure. Lysozyme and BSA were fast separated within 10 min using this affinity column. The column was also used for the purification of lysozyme from chicken egg white.