title: Poly(hydroxyethyl methacrylate) based affinity cryogel for plasmid DNA purification.
abstract: 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.
authors.affiliation: Department of Biology, Hacettepe University, Ankara, Turkey.
authors.forename: Işık
authors.initials: I
authors.lastname: Perçin
authors.forename: Emel
authors.initials: E
authors.lastname: Sağlar
authors.forename: Handan
authors.initials: H
authors.lastname: Yavuz
authors.forename: Erol
authors.initials: E
authors.lastname: Aksöz
authors.forename: Adil
authors.initials: A
authors.lastname: Denizli
mesh_terms: D000327:Adsorption; D002846:Chromatography, Affinity / Q000379:methods*; D059985:Cryogels; D004269:DNA, Bacterial / Q000302:isolation & purification*; D006639:Histidine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D020100:Hydrogels / Q000737:chemistry*; D008689:Methacrylates / Q000138:chemical synthesis* / Q000737:chemistry; D010957:Plasmids / Q000235:genetics*; D011102:Polyhydroxyethyl Methacrylate / Q000737:chemistry*
chemical_list: D059985:Cryogels; D004269:DNA, Bacterial; D020100:Hydrogels; D008689:Methacrylates; C491950:N-methacryloyl-L-histidine methyl ester; D011102:Polyhydroxyethyl Methacrylate; D006639:Histidine