Dye affinity cryogels for plasmid DNA purification

Mater Sci Eng C Mater Biol Appl. 2015 Nov 1;56:318-24. doi: 10.1016/j.msec.2015.06.041. Epub 2015 Jun 23.

Abstract

The aim of this study is to prepare megaporous dye-affinity cryogel discs for the purification of plasmid DNA (pDNA) from bacterial lysate. Poly(hydroxyethyl methacrylate) [PHEMA] cryogel discs were produced by free radical polymerization initiated by N,N,N',N'-tetramethylene diamine (TEMED) and ammonium persulfate (APS) redox pair in an ice bath. Cibacron Blue F3GA was used as an affinity ligand (loading amount: 68.9μmol/g polymer). The amount of pDNA adsorbed onto the PHEMA-Cibacron Blue F3GA cryogel discs first increased and then reached a plateau value (i.e., 32.5mg/g cryogel) at 3.0mg/mL pDNA concentration. Compared with the PHEMA cryogel (0.11mg/g cryogel), the pDNA adsorption capacity of the PHEMA-Cibacron Blue F3GA cryogel (32.4mg/g polymer) was improved significantly due to the Cibacron Blue 3GA immobilization onto the polymeric matrix. pDNA adsorption amount decreased from 11.7mg/g to 1.1mg/g with the increasing of NaCl concentration. The maximum pDNA adsorption was achieved at 4°C. The overall recovery of pDNA was calculated as 90%. The PHEMA-Cibacron Blue F3GA cryogel discs could be used five times without decreasing the pDNA adsorption capacity significantly. The results show that the PHEMA-Cibacron Blue F3GA cryogel discs promise high selectivity for pDNA.

Keywords: Affinity adsorption; Cibacron Blue F3GA; Cryogels; DNA purification; PHEMA.

MeSH terms

  • Cryogels / chemistry*
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / isolation & purification*
  • Escherichia coli / chemistry*
  • Plasmids / chemistry
  • Plasmids / isolation & purification*
  • Polyhydroxyethyl Methacrylate / chemistry*
  • Triazines / chemistry*

Substances

  • Cryogels
  • DNA, Bacterial
  • Triazines
  • Polyhydroxyethyl Methacrylate
  • Cibacron Blue F 3GA