Dye affinity cryogels for plasmid DNA purification
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Dosyalar
Tarih
2015
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Elsevier
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
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: 68.9 mu mol/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., 32.5 mg/g cryogel) at 3.0 mg/mL pDNA concentration. Compared with the PHEMA cryogel (0.11 mg/g cryogel), the pDNA adsorption capacity of the PHEMA-Cibacron Blue F3GA cryogel (32.4 mg/g polymer) was improved significantly due to the Cibacron Blue 3GA immobilization onto the polymeric matrix. pDNA adsorption amount decreased from 11.7 mg/g to 1.1 mg/g with the increasing of NaCl concentration. The maximum pDNA adsorption was achieved at 4 degrees 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. (C) 2015 Elsevier B.V. All rights reserved.
Açıklama
Anahtar Kelimeler
PHEMA, Cryogels, Cibacron Blue F3GA, DNA purification, Affinity adsorption
Kaynak
Materials Science & Engineering C-Materials For Biological Applications
WoS Q Değeri
Q2
Scopus Q Değeri
Cilt
56
