New monolith technology for automated anion-exchange purification of nucleic acids

Thayer, James R.; Flook, Kelly; Woodruff, Andy; Rao, S. Narasimha; Pohl, Christopher A.

doi:10.1016/j.jchromb.2010.01.030

Cited by 34 publications

(16 citation statements)

References 37 publications

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“…Recently, chromatographic techniques such as RP (toxic), LC, ion‐pairing RP (toxic), gel filtration and anion‐exchange , and affinity techniques such as base‐pairing , affinity tags and amino acids‐RNA have been used to isolate, separate, and purify RNA and DNA. However, these techniques have some drawbacks such as the need for toxic chemicals, scale‐up problems, time consuming, denaturation, misfolding impurity, instability, or the need for high salt concentration.…”

Section: Introductionmentioning

confidence: 99%

PolyGuanine methacrylate cryogels for ribonucleic acid purification

Köse

Uzun

2016

J of Separation Science

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The isolation and purification of ribonucleic acid have attracted attention recently for the understanding of the functions in detail because of the necessity for the treatment of genetic diseases. In this study, guanine-incorporated polymeric cryogels were developed to obtain highly purified ribonucleic acid. The satisfactory purification performance was achieved with the guanine-incorporated poly (2-hydroxyethyl methacrylate-guanine methacrylate) cryogels. The most crucial advantages to use guanine as a functional monomer are to obtain a real natural interaction between guanine on the polymeric material and cytosine on the ribonucleic acid. Moreover, using cryogel with a highly porous structure and high swelling ratio provide advantages of getting more water within the structure to get more analyte to interact. The characterization of cryogels has proved the success of the synthesis and the perfect natural interaction to be taken place between the ligand (guanine methacrylate) and the cytosine in the ribonucleic acid molecules. Although the pores within the structure of cryogels are small, they provide efficient and fast adsorption. The chromatographic separation performance was investigated for different conditions (pH, temperature etc.). The desorption ratio and reusability were also analyzed at the end of the five adsorption-desorption cycles with no significant changes.

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Section: Introductionmentioning

confidence: 99%

PolyGuanine methacrylate cryogels for ribonucleic acid purification

Köse

Uzun

2016

J of Separation Science

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“…To date, there are no reports of resolution between peaks differing by only one base for this class of oligonucleotides. [66]. The stationary phase uses porous ionexchange "nanobeads" that are attached to monolithic polymers on a 4.6 × 50 mm column.…”

Section: Ion-exchange Chromatographymentioning

confidence: 99%

“…The new columns are more stable at higher temperature and pH and exhibit a much improved column lifetime. A monolithic stationary phase with porous ion-exchange "nanobeads" has recently been introduced [66]. Both the methacrylate and monolithic columns are now available in custom 1 mm and capillary sizes as companies move toward supporting nano chromatography applications that have already been so successful in proteomics.…”

Section: Columns and Stationary Phases For Ion-exchange Chromatographymentioning

confidence: 99%

Chromatographic methods for the determination of therapeutic oligonucleotides

McGinnis

Chen

Bartlett

2012

Journal of Chromatography B

112

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“…This type of stationary phase is composed of a macroporous continuous (interconnected) support structure typically made from organic polymers 41 or silica precursors 42. Macroporous polymer monoliths have shown great potential for gradient separations of biomolecules, including peptides 43–45, nucleic acids 46, and intact proteins 47. Silica monoliths are characterised by a macroporous bed containing mesoporous silica skeletons and have been more successfully utilised for small molecule separations 48, 49.…”

Section: New Stationary Phase Morphologies: Possibilities and Limitmentioning

confidence: 99%

Kinetic performance optimisation for liquid chromatography: Principles and practice

Causon

Broeckhoven

Hilder

et al. 2011

J of Separation Science

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This HPLC tutorial focuses on the preparation and use of kinetic plots to characterise the performance in isocratic and gradient LC. This graphical approach allows the selection of columns (i.e. optimum particle size and column length) and LC conditions (operating pressure and temperature) to generate a specific number of plates or peak capacity in the shortest possible analysis time. Instrument aspects including the influence of extra-column effects (maximum allowable system volume) and thermal operating conditions (oven type) on performance are discussed. In addition, the performance characteristics of porous-shell particle-packed columns and monolithic stationary phases are presented and the potential of future column designs is discussed.

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New monolith technology for automated anion-exchange purification of nucleic acids

Cited by 34 publications

References 37 publications

PolyGuanine methacrylate cryogels for ribonucleic acid purification

PolyGuanine methacrylate cryogels for ribonucleic acid purification

Chromatographic methods for the determination of therapeutic oligonucleotides

Kinetic performance optimisation for liquid chromatography: Principles and practice

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