Design and application of a newly generated bio/synthetic cryogel column for DNA capturing
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Dosyalar
Tarih
2020
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Springer Science and Business Media Deutschland GmbH
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
The need for high amounts of plasmid (p) and genomic (g) DNA separation is increasing day by day for gene therapy, DNA vaccination, etc. Here, extracellular polymeric substances (EPS) used as a binding agent for Fe3+ ions, which have high affinity to DNA molecules, were mixed with 2-hydroxyethyl methacrylate (HEMA) to obtain EPS–PHEMA-blended cryogel column for separation of gDNA and pDNA. The obtained Fe3+–EPS–PHEMA column was analysed by scanning electron microscopy and swelling test measurements. Effect of pH, initial DNA concentration, ionic strength, flow rate and temperature on separation of DNA was studied. Adsorbed gDNA from aqueous solution was 39.7 mg/g in acetate buffer at pH 6 with the initial DNA concentration of 2 mg/mL and the flow rate of 0.5 mL/min at 25 °C. A260/A280 ratio and adsorption capacity of Fe3+–EPS–PHEMA composite cryogels for pDNA extracted from bacterial lysates were found to be 1.85 and 38.6, respectively. Fe3+–EPS–PHEMA composite cryogels have a monolayer adsorption according to the Langmuir model. The Temkin model was utilized to characterize adsorption energetically. The prepared sorbent was tested for some kinetic parameters too. It was seen that the second-order mechanism arguing chemisorption is dominant for DNA adsorption. The reuse of matrix was tested over 30 times without any deformation in shape and adsorption capacity losing.
Açıklama
Anahtar Kelimeler
Composite Cryogels, DNA Separation, Extracellular Polymeric Substances, IMAC
Kaynak
Polymer Bulletin
WoS Q Değeri
Q2
Scopus Q Değeri
Q1
Cilt
-
Sayı
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