Plasmid DNA purification using a multimodal chromatography resin

Abstract

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.

Comparative DNA isolation behaviours of silica and polymer based sorbents in batch fashion: monodisperse silica microspheres with bimodal pore size distribution as a new sorbent for DNA isolation

“…Methacrylate based monolith was prepared by bearing diethylaminoethyl (DEAE) and butyl groups and used as a sorbent for selective extraction of DNA [20]. Matos et al used a chromatography medium (resin) to isolate plasmid DNA [21]. The magnetic hybrid nanocomposites were synthesized to perform simple and fast DNA separation [22].…”

Screening of l‐histidine‐based ligands to modify monolithic supports and selectively purify the supercoiled plasmid DNA isoform

“…This behavior is also confirmed in Table , because the affinity differences between oc and sc pVAX1‐ LacZ isoforms in the 1‐benzyl‐ l ‐histidine (5.62 × 10 −8 ± 0.689 × 10 −8 M for oc and 6.03 × 10 −9 ± 0.238 × 10 −9 M for sc isoforms) are higher than the affinity differences in the l ‐histidine (8.18 × 10 −8 ± 0.752 × 10 −8 M for oc and 5.79 × 10 −8 ± 0.46 × 10 −8 M for sc isoforms). This result suggests that beyond the aforementioned interactions established between the imidazole ring and the sc pDNA isoform, additional interactions can be involved because of the hydrophobic character of the benzyl aromatic ring (Matos et al ., ).…”

Improved ionic-liquid-functionalized macroporous supports able to purify nucleic acids in one step

“…3 B. This chromatographic behavior where more than one form of non-covalent chemical interactions occurs between the stationary phase and the target molecules is typical of multimodal ligands [ 44 ], where two main types of interactions, namely hydrophobic and electrostatic interactions, occur between RNA and MP-SilPrMImCl. These interactions are expected given the IL chemical structure, which has an aromatic ring and an aliphatic moiety, and it is positively charged.…”