Çimen, DuyguYılmaz, FatmaPerçin, IşıkTürkmen, DenizDenizli, Adil2024-09-252024-09-2520150928-49311873-0191https://doi.org/10.1016/j.msec.2015.06.041https://hdl.handle.net/20.500.12491/13912The 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.eninfo:eu-repo/semantics/openAccessPHEMACryogelsCibacron Blue F3GADNA purificationAffinity adsorptionArticle10.1016/j.msec.2015.06.0415631832426249596WOS:000359873900038Q2