Department of Biology, Hacettepe University, Ankara, Turkey.
Int J Biol Macromol. 2011 May 1;48(4):577-82. doi: 10.1016/j.ijbiomac.2011.01.022. Epub 2011 Feb 2.
The aim of this study is to prepare supermacroporous pseudospecific cryogel which can be used for the purification of plasmid DNA (pDNA) from bacterial lysate. N-methacryloyl-(l)-histidine methyl ester (MAH) was chosen as the pseudospecific ligand and/or comonomer. Poly(hydroxyethyl methacrylate-N-methacryloyl-(l)-histidine methyl ester) [PHEMAH] cryogel was produced by free radical polymerization initiated by N,N,N',N'-tetramethylene diamine (TEMED) and ammonium persulfate (APS) pair in an ice bath. Compared with the PHEMA cryogel (50 μg/g polymer), the pDNA adsorption capacity of the PHEMAH cryogel (13,350 μg/g polymer) was improved significantly due to the MAH incorporation into the polymeric matrix. The amount of pDNA bound onto the PHEMAH cryogel disks first increased and then reached a saturation value (i.e., 13,350μg/g) at around 300 μg/ml pDNA concentration. pDNA adsorption amount decreased from 1137 μg/g to 160 μg/g with the increasing NaCl concentration. The maximum pDNA adsorption was achieved at 25 °C. The overall recovery of pDNA was calculated as 90%. The PHEMAH cryogel could be used 3 times without decreasing the pDNA adsorption capacity significantly. The results indicate that the PHEMAH cryogel disks promise high selectivity for pDNA.
本研究旨在制备可用于从细菌裂解物中纯化质粒 DNA（pDNA）的超大孔假特异性 cryogel。选择 N-丙烯酰基-(l)-组氨酸甲酯（MAH）作为假特异性配体和/或共聚单体。聚（羟乙基甲基丙烯酸酯-N-丙烯酰基-(l)-组氨酸甲酯）[PHEMAH] cryogel 通过在冰浴中由 N,N,N',N'-四亚甲基二胺（TEMED）和过硫酸铵（APS）引发的自由基聚合产生。与 PHEMA cryogel（50 μg/g 聚合物）相比，由于 MAH 掺入聚合物基质，PHEMAH cryogel 的 pDNA 吸附能力（13,350 μg/g 聚合物）显著提高。PHEMAH cryogel 盘上结合的 pDNA 量首先增加，然后在约 300μg/ml pDNA 浓度时达到饱和值（即 13,350μg/g）。随着 NaCl 浓度的增加，pDNA 吸附量从 1137μg/g 下降到 160μg/g。pDNA 的最大吸附在 25°C 时达到。pDNA 的总体回收率计算为 90%。PHEMAH cryogel 可以使用 3 次而不会显著降低 pDNA 吸附能力。结果表明，PHEMAH cryogel 盘对 pDNA 具有高选择性。
Zotero 插件