Aromatic ligands for plasmid deoxyribonucleic acid chromatographic analysis and purification: An overview

Caramelo-Nunes, C.; Almeida, Paulo; Marcos, João Carlos; Tomaz, Cândida T.

doi:10.1016/j.chroma.2013.12.057

Cited by 10 publications

(7 citation statements)

References 161 publications

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“…The elemental analysis results given in Table 1 show no significant differences in the carbon and hydrogen content ( wt %) for both matrices, namely MP and MP-SilPrMImCl supports; however, nitrogen (2.03 wt %) was detected in the final modified support, as well as in MP-MIm support, confirming the presence of the imidazolium ring. Based on the nitrogen content provided by elemental analysis, the ligand density [ 41 ] of MP-SilPrMImCl (given in Table 1 ) corresponds to 0.725 mmol of IL per gram of support, a higher value than that present in supports modified with common ligands used in the separation of nucleic acids (naphthalene tripodal: 0.32 mmol/g sepharose CL-6B [ 41 ], and 3,8-diamino-6-phenylphenathridine: 0.15 mmol/g derivatized sepharose [ 42 ]). Taking all characterization results into consideration, it is shown that the IL is efficiently and covalently bound to the support.…”

Section: Resultsmentioning

confidence: 99%

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

Neves

Pereira

Pedro

et al. 2020

Materials Today Bio

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Section: Resultsmentioning

confidence: 99%

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

Neves

Pereira

Pedro

et al. 2020

Materials Today Bio

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“…At these pH conditions, the histidine‐based ligands can interact with the phosphate groups and the bases of nucleic acids. Because the pDNA isoform presents higher charge density per surface area and more exposed aromatic bases due to the torsion associated to supercoiling , the accentuated increase of RT with both columns is understandable. These results suggest an increased selectivity for sc pDNA by using acidic pH.…”

Section: Resultsmentioning

confidence: 99%

Chromatographic HPV‐16 E6/E7 plasmid vaccine purification employing L‐histidine and 1‐benzyl‐L‐histidine affinity ligands

et al. 2017

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Affinity chromatography based on amino acids as interacting ligands was already indicated as an alternative compared to ion exchange or hydrophobic interaction for plasmid DNA purification. Understanding the recognition mechanisms occurring between histidine-based ligands and nucleic acids enables more efficient purification of a DNA vaccine, as the binding and elution conditions can be adjusted in order to enhance the purification performance. Decreasing pH to slightly acidic conditions increases the positive charge of histidine ligand, what influences the type of interaction between chromatographic support and analytes. This was proven in this work, where hydrophobic effects established in the presence of ammonium sulfate were affected at pH 5.0 in comparison to pH 8.0, while electrostatic and cation-π interactions were intensified. Histidine ligand at pH 5.0 interacts with phosphate groups or aromatic rings of plasmid DNA. Due to different responses of RNA and pDNA on mobile phase changes, the elution order between RNA and pDNA was changed with mobile phase pH decrease from 8.0 to 5.0. The phenomenon was more evident with L-histidine ligand due to more hydrophilic character, leading to an improved selectivity of L-histidine-modified chromatographic monolith, allowing the product recovery with 99% of purity (RNA removal). With the 1-benzyl- L-histidine ligand, stronger and less selective interactions with the nucleic acids were observed due to the additional hydrophobicity associated with the phenyl aromatic ring. Optimization of sample displacement chromatography parameters (especially (NH ) SO concentration) at slightly acidic pH enabled excellent isolation of pDNA, by the removal of RNA in a negative mode, with binding capacities above 1.5 mg pDNA per mL of chromatographic support.

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“…This behavior suggests that besides the predominant action of hydrophobic interactions, owing to the salt concentration used, other interactions can be present, affecting the plasmid retention. Given that the p K a of l ‐histidine side chain is 6.5, acidic pH environment can favor the protonation of imidazole ring, which enables the additional involvement of ring stacking interactions and cation π interactions with plasmid aromatic bases (Caramelo‐Nunes et al ., ), especially with guanine (Luscombe et al ., ; Sousa et al ., ).…”

Section: Resultsmentioning

confidence: 99%

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

Amorim

Sousa

Queiroz

et al. 2015

J of Molecular Recognition

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The growing demand of pharmaceutical-grade plasmid DNA (pDNA) suitable for biotherapeutic applications fostered the development of new purification strategies. The surface plasmon resonance technique was employed for a fast binding screening of l-histidine and its derivatives, 1-benzyl-L-histidine and 1-methyl-L-histidine, as potential ligands for the biorecognition of three plasmids with different sizes (6.05, 8.70, and 14 kbp). The binding analysis was performed with different isoforms of each plasmid (supercoiled, open circular, and linear) separately. The results revealed that the overall affinity of plasmids to l-histidine and its derivatives was high (KD > 10(-8) M), and the highest affinity was found for human papillomavirus 16 E6/E7 (K(D) = 1.1 × 10(-10) M and KD = 3.34 × 10(-10) M for open circular and linear plasmid isoforms, respectively). L-Histidine and 1-benzyl-L-histidine were immobilized on monolithic matrices. Chromatographic studies of L-histidine and 1-benzyl-L-histidine monoliths were also performed with the aforementioned samples. In general, the supercoiled isoform had strong interactions with both supports. The separation of plasmid isoforms was achieved by decreasing the ammonium sulfate concentration in the eluent, in both supports, but a lower salt concentration was required in the 1-benzyl-L-histidine monolith because of stronger interactions promoted with pDNA. The efficiency of plasmid isoforms separation remained unchanged with flow rate variations. The binding capacity for pDNA achieved with the l-histidine monolith was 29-fold higher than that obtained with conventional L-histidine agarose. Overall, the combination of either L-histidine or its derivatives with monolithic supports can be a promising strategy to purify the supercoiled isoform from different plasmids with suitable purity degree for pharmaceutical applications.

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Aromatic ligands for plasmid deoxyribonucleic acid chromatographic analysis and purification: An overview

Cited by 10 publications

References 161 publications

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

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

Chromatographic HPV‐16 E6/E7 plasmid vaccine purification employing L‐histidine and 1‐benzyl‐L‐histidine affinity ligands

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

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