A multimodal histamine ligand for chromatographic purification of plasmid DNA

Sample displacement chromatography of plasmid DNA isoforms

“…The achieved pyridine ligand density on the chromatographic support was 1.0 mmol/mL, determined from elemelemental analysis. Histamine was immobilised on CIMac CDI monolith leading to the neutral carbamate bond, as it is described thoroughly in reference [19].…”

“…Pharmaceutical-grade pDNA requires high homogeneity of supercoiled (sc) form, and chromatographic separation of sc from open coiled (oc) or linear isoforms on preparative scale is extremely challenging due to their structural similarity [19]. DC/SDC approaches have not yet been investigated for the separation of pDNA isoforms, and the main purpose of the present work was to develop a method for separation of sc pDNA from other pDNA isoforms using SDC on monolithic chromatographic supports, that would enable high resolution, high capacity and scalable pDNA purification methods in the future.…”

Plasmid DNA purification using a multimodal chromatography resin

“…However, only partial separation could be achieved, and when a linear NaCl gradient was explored, an sc pDNA purity of 70% was obtained in an optimized system. The Capto Adhere column can therefore not compete with other more specific resins in this aspect, such as MMC histamine (e.g., Černigoj et al ., ). More crude and complex samples were subsequently tested, and the chromatographic profile of pDNA elution from a clarified E. coli lysate is presented in Figure .…”

“…Compared with the traditional mono‐modal ligands, a higher selectivity can often be achieved with these ligands. Several amino acid‐based ligands have been utilized for pDNA separations, and these resins have MMC‐like behavior (e.g., Černigoj et al ., ). In this study, we have examined Capto Adhere , which is a recent MMC support material (Figure ) based on HIC and AEX type of interactions (Chen et al ., ).…”

Plasmid production and purification: An integrated experiment‐based biochemistry and biotechnology laboratory course

“…The imidazole group of L‐histidine has some unique features that may result in several multiple non‐covalent interactions with sc pVAX1‐ LacZ bases . In fact, despite the prevalence of hydrophobic interactions, other multiple noncovalent interactions such as electrostatic interactions, hydrogen bonding and π–π stacking can also provide resources selectivity, as previously reported to other ligands . In particular, histamine was used as an immobilized chromatographic ligand for the purification of sc pDNA by establishing mainly hydrophobic interactions, although electrostatic interactions can also play an important role in the recognition of pDNA .…”