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Dye affinity cryogels for plasmid DNA purification
- Source :
- Materials Science and Engineering: C. 56:318-324
- Publication Year :
- 2015
- Publisher :
- Elsevier BV, 2015.
-
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.
- Subjects :
- DNA, Bacterial
chemistry.chemical_classification
Materials science
Chromatography
Triazines
Ligand
Radical polymerization
Bioengineering
Polymer
(Hydroxyethyl)methacrylate
Redox
Biomaterials
chemistry.chemical_compound
Adsorption
chemistry
Mechanics of Materials
Diamine
Escherichia coli
Ammonium persulfate
Cryogels
Polyhydroxyethyl Methacrylate
Plasmids
Subjects
Details
- ISSN :
- 09284931
- Volume :
- 56
- Database :
- OpenAIRE
- Journal :
- Materials Science and Engineering: C
- Accession number :
- edsair.doi.dedup.....464b7257c7fdac5695a07d3d17475871