PREPARATION OF A BIPOROUS POLYMERIC ADSORBENT FOR RAPID CHROMATOGRAPHIC PURIFICATION OF PLASMID DNA
PREPARATION OF A BIPOROUS POLYMERIC ADSORBENT FOR RAPID CHROMATOGRAPHIC PURIFICATION OF PLASMID DNA
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用于快速层析纯化质粒DNA的双孔聚合物吸附剂的制备
DOI：
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发表时间：
2006
期刊：
影响因子：
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通讯作者：
Sun Guoyong
中科院分区：
文献类型：
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作者：
Sun Yan;Sun Guoyong
本文报道了一种用于高速流动色谱的双孔聚合物固定相的制备方法，用200 g/L碳酸钙悬浮液和有机溶剂（环己醇和十二烷醇的混合物）作为致孔剂，以甲基丙烯酸缩水甘油酯和二甲基丙烯酸乙二醇酯为单体，采用（W/O）/（W/O）（W/O W乳化，用二乙胺对基质进行改性，制备了阴离子交换剂（DEA-B），发现其具有两类孔（微孔和超孔），而O/W乳化法制备的微孔微球（DEA-M）中只有微孔，DEA-B中的两族孔分布在50~ 100 μ m之间DEA-B的水化密度和粒径分别为1.08 g/mL和46.3 μm，与DEA-M相近DEAB柱在移动的流速为150 cm/h的条件下，可以纯化5.4 kb质粒DNA，而DEA-M柱在相同条件下不能纯化质粒DNA。DEA-B柱在1500 cm/h的流速下可获得高效液相色谱纯质粒DNA，表明DEA-B柱可用于质粒DNA的快速层析纯化。
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.