Analysis of oligonucleotides by liquid chromatography with alkylamide stationary phase

Buszewski, Bogusław; Safaei, Zahra; Studzińska, Sylwia

doi:10.1515/chem-2015-0141

Cited by 8 publications

(6 citation statements)

References 18 publications

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“…The alkylamide stationary phase was also applied to OGN studies in RP HPLC and IPC [ 57 , 73 ]. Compared to C18, the retention of OGNs was lower [ 56 , 63 ].…”

Section: Application Of Adsorbents With Different Modifications For S...mentioning

confidence: 99%

“…The polar heteroatoms implemented in alkyl chains (e.g., in the amide group) increase the polarity of the stationary phase and, at the same time, reduced the retention of OGNs ( Figure 4 ) [ 53 , 57 , 64 ]. It was also shown that OGN retention increases with the decrease of the mobile phase pH in RP mode, indicating the critical role of surface charge change [ 73 ]. The advantage of the alkylamide stationary phase is that the application of RP HPLC allowed for the successful separation of sequence isomers at acidic pH containing a high concentration of buffer.…”

Section: Application Of Adsorbents With Different Modifications For S...mentioning

confidence: 99%

“…The advantage of the alkylamide stationary phase is that the application of RP HPLC allowed for the successful separation of sequence isomers at acidic pH containing a high concentration of buffer. Contrastingly, the time needed for separation was long, and peak asymmetry was significant due to the amide and aminopropyl groups’ presence [ 73 ]. Greater OGN peak symmetry was obtained for a similar stationary phase when UHPLC was used in ion pair mode [ 63 , 74 ].…”

Section: Application Of Adsorbents With Different Modifications For S...mentioning

confidence: 99%

See 2 more Smart Citations

Oligonucleotides Isolation and Separation—A Review on Adsorbent Selection

Studzińska

Nuckowski

Buszewski

2022

IJMS

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Oligonucleotides have many important applications, including as primers in polymerase chain reactions and probes for DNA sequencing. They are proposed as a diagnostic and prognostic tool for various diseases and therapeutics in antisense therapy. Accordingly, it is necessary to develop liquid chromatography and solid phase extraction methods to separate oligonucleotides and isolate them from biological samples. Many reviews have been written about the determination of these compounds using the separation technique or sample preparation for their isolation. However, presumably, there are no articles that critically review the adsorbents used in liquid chromatography or solid phase extraction. The present publication reviews the literature from the last twenty years related to supports (silica, polymers, magnetic nanoparticles) and their modifications. The discussed issues concern reversed phase (alkyl, aromatic, cholesterol, mixed ligands), ion-exchange (strong and weak ones), polar (silica, polyhydroxy, amide, zwitterionic), and oligonucleotide-based adsorbents.

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“…The alkylamide stationary phase was also applied to OGN studies in RP HPLC and IPC [ 57 , 73 ]. Compared to C18, the retention of OGNs was lower [ 56 , 63 ].…”

Section: Application Of Adsorbents With Different Modifications For S...mentioning

confidence: 99%

Section: Application Of Adsorbents With Different Modifications For S...mentioning

confidence: 99%

Section: Application Of Adsorbents With Different Modifications For S...mentioning

confidence: 99%

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Oligonucleotides Isolation and Separation—A Review on Adsorbent Selection

Studzińska

Nuckowski

Buszewski

2022

IJMS

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“…24 In an HPLC column, the surface of the adsorbents is modified by various functional groups, such as amino propyl 25 and amide groups. 26 Although many adsorbents have been developed for effective separation and purification, efficient separation conditions (pH, organic solvent ratio, and functional group) must be investigated through trial and error. Therefore, a theoretical explanation of the DNA separation mechanism is essential for further chromatographic efficiency.…”

Section: Introductionmentioning

confidence: 99%

Distribution Behavior of Single-Stranded DNA Molecules in an Amino-Group-Functionalized Silica Microparticle

et al. 2022

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In this study, we investigated the distribution behavior of single-stranded DNA molecules with 20 bases in silica particles (particle size: ∼30 μm) using confocal fluorescence microspectroscopy. The distribution kinetics was investigated under various conditions, such as the type of base (adenine, thymine, guanine, and cytosine), pore size of the particle (30 and 50 nm), and salt concentration (100, 200, and 500 mM), which changed the distribution behavior. At high salt concentrations, we observed sigmoidal kinetic behavior, which does not occur in the general distribution of small organic molecules but is often observed in protein aggregation and nuclear growth. An analytical model based on DNA aggregation explained the sigmoidal distribution behavior well, and this model also worked well when the number of DNA molecules involved in DNA aggregation was greater than two. The intraparticle diffusion of DNA molecules was analyzed using the pore and surface diffusion model. As a result, the intraparticle diffusion of DNA aggregates mainly occurs according to surface diffusion, and the surface diffusion coefficient has the same value ((2.4−6.7) × 10 −9 cm 2 s −1 ) independent of the pore size and type of base.

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“…pH of the mobile phase also plays an important role to control retention and selectivity. For example, in the case of alkylamide stationary phases, as reported in Ref [110],. when pH is buffered at 6 or 7, the negative charges on the stationary phase are not efficiently shielded by the salts.…”

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confidence: 99%

Oligonucleotides: Current Trends and Innovative Applications in the Synthesis, Characterization, and Purification

Catani

Luca

Alcântara

et al. 2020

Biotechnology Journal

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Oligonucleotides (ONs) are gaining increasing importance as a promising novel class of biopharmaceuticals. Thanks to their fundamental role in gene regulation, they can be used to develop custom-made drugs (also called N-to-1) able to act on the gene expression at pretranslational level. With recent approvals of ON-based therapeutics by FDA, a growing demand for high-quality chemically-modified ONs is emerging and their market is expected to impressively prosper in the very next future. To satisfy this growing market demand, a scalable and economically sustainable ON production is needed. In this paper, the state of the art of the whole ON production process is illustrated with the aim of highlighting the most promising routes towards the auspicated market-size production. In particular, the most recent advancements in both the upstream stage, mainly based on solid-phase synthesis and recombinant technology, and the downstream one, focusing on chromatographic techniques, are reviewed. Since ON production is projected to expand to the large scale, automatized multi-column technologies will reasonably be required soon to replace the current ones based on batch single-column operations. Cutting edge purification solutions, based on continuous chromatography, will be thus presented in the last part of this review.

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Analysis of oligonucleotides by liquid chromatography with alkylamide stationary phase

Cited by 8 publications

References 18 publications

Oligonucleotides Isolation and Separation—A Review on Adsorbent Selection

Oligonucleotides Isolation and Separation—A Review on Adsorbent Selection

Distribution Behavior of Single-Stranded DNA Molecules in an Amino-Group-Functionalized Silica Microparticle

Oligonucleotides: Current Trends and Innovative Applications in the Synthesis, Characterization, and Purification

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