Poly(hydroxyethyl methacrylate) based affinity cryogel for plasmid DNA purification


Percin I. , Saglar E. , Yavuz H. , Aksoz E., DENİZLİ A.

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, vol.48, no.4, pp.577-582, 2011 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 48 Issue: 4
  • Publication Date: 2011
  • Doi Number: 10.1016/j.ijbiomac.2011.01.022
  • Title of Journal : INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
  • Page Numbers: pp.577-582

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) [PHEMAI-1] 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 mu g/g polymer), the pDNA adsorption capacity of the PHEMAH cryogel (13,350 mu g/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., 13,350 mu g/g) at around 300 mu g/ml pDNA concentration. pDNA adsorption amount decreased from 1137 mu g/g to 160 mu g/g with the increasing NaCl concentration. The maximum pDNA adsorption was achieved at 25 degrees 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. (C) 2011 Elsevier B.V. All rights reserved.