PREPARATION OF A BIPOROUS POLYMERIC ADSORBENT FOR RAPID CHROMATOGRAPHIC PURIFICATION OF PLASMID DNA

Abstract

Preparation of a biporous polymeric stationary phase for high-speed flowthrough chromatography was reported.Using calcium carbonate suspension(200 g/L) and organic solvent(mixture of cyclohexanol and dodecanol) as the porogenic agents,a novel biporous matrix has been prepared by ultraviolet photo-polymerizing monomers of glycidyl methacrylate and ethylene glycol dimethacrylate by(W/O)/W emulsification.The matrix was modified with diethylamine to create an anion exchanger(DEA-B).It was found that there was two families of pores(micropores and superpores) in DEA-B,but there was only micropores in the microporous beads(DEA-M) prepared by O/W emulsification.The two families of pores in DEA-B were in the range of 50～200 nm and 500～5000 nm,respectively,while the pores of DEA-M were mainly in the range of 50～200 nm.The hydrated density and particle size of DEA-B were 1.08 g/mL and 46.3 μm,respectively,similar to those of DEA-M(1.10 g/mL and 45.2 μm).Operated at a mobile phase velocity of 150 cm/h,the DEAB column could be used to purify a 5.4 kb plasmid DNA,but the DEA-M column failed to do that at the same condition.Then,HPLC pure plasmid DNA was were obtained with the DEA-B column operated at 1500 cm/h.The results indicated that DEA-B column was promising for high-speed chromatography purification of plasmid DNA.