Plasmid DNA purification using a multimodal Chromatography resin

Research output: Contribution to journalArticle

Standard

Plasmid DNA purification using a multimodal Chromatography resin. / Matos, Tiago; Queiroz, João A.; Bülow, Leif.

In: Journal of Molecular Recognition, Vol. 27, No. 4, 2014, p. 184-189.

Research output: Contribution to journalArticle

Harvard

APA

CBE

MLA

Vancouver

Author

Matos, Tiago ; Queiroz, João A. ; Bülow, Leif. / Plasmid DNA purification using a multimodal Chromatography resin. In: Journal of Molecular Recognition. 2014 ; Vol. 27, No. 4. pp. 184-189.

RIS

TY - JOUR

T1 - Plasmid DNA purification using a multimodal Chromatography resin

AU - Matos, Tiago

AU - Queiroz, João A.

AU - Bülow, Leif

PY - 2014

Y1 - 2014

N2 - Abstract in Undetermined Multimodal chromatography is widely used for isolation of proteins because it often results in improved selectivity compared to conventional separation resins. The binding potential and chromatographic behavior of plasmid DNA have here been examined on a Capto Adhere resin. Capto Adhere is a recent multimodal chromatography material allowing molecular recognition between the ligand and target molecule, which is based on combined ionic and aromatic interactions. Capto Adhere proved to offer a very strong binding of nucleic acids. This property could be used to isolate plasmid DNA from a crude Escherichia coli extract. Using a stepwise NaCl gradient, pure plasmid DNA could be obtained without protein and endotoxin contaminations. The RNA fraction bound most strongly to the resin and could be eluted only at very high salt concentrations (2.0 M NaCl). The chromatographic separation behavior was very robust between pH values 6 and 9, and the dynamic binding capacity was estimated to 60 µg/ml resin.

AB - Abstract in Undetermined Multimodal chromatography is widely used for isolation of proteins because it often results in improved selectivity compared to conventional separation resins. The binding potential and chromatographic behavior of plasmid DNA have here been examined on a Capto Adhere resin. Capto Adhere is a recent multimodal chromatography material allowing molecular recognition between the ligand and target molecule, which is based on combined ionic and aromatic interactions. Capto Adhere proved to offer a very strong binding of nucleic acids. This property could be used to isolate plasmid DNA from a crude Escherichia coli extract. Using a stepwise NaCl gradient, pure plasmid DNA could be obtained without protein and endotoxin contaminations. The RNA fraction bound most strongly to the resin and could be eluted only at very high salt concentrations (2.0 M NaCl). The chromatographic separation behavior was very robust between pH values 6 and 9, and the dynamic binding capacity was estimated to 60 µg/ml resin.

U2 - 10.1002/jmr.2349

DO - 10.1002/jmr.2349

M3 - Article

VL - 27

SP - 184

EP - 189

JO - Journal of Molecular Recognition

T2 - Journal of Molecular Recognition

JF - Journal of Molecular Recognition

SN - 1099-1352

IS - 4

ER -