Liquid chromatographic separation of oligonucleotides

Biba, Mirlinda; Foley, Joe P.; Welch, Christopher J.

doi:10.1016/b978-0-12-805392-8.00006-2

Cited by 9 publications

(11 citation statements)

References 81 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The pKa of the phosphodiester linkage is 2, which means that the oligonucleotides contain one negative charge for every phosphodiester linkage in an aqueous solution above pH 4. Therefore, traditional RP-LC conditions tend not to work well for the separation of oligonucleotides, and ion-exchange chromatography techniques or ion-paired RPLC need to be considered [29]. …”

Section: Separation and Purification Of Oligonucleotidesmentioning

confidence: 99%

Recent Methods for Purification and Structure Determination of Oligonucleotides

Zhang

Wang

et al. 2016

IJMS

View full text Add to dashboard Cite

Aptamers are single-stranded DNA or RNA oligonucleotides that can interact with target molecules through specific three-dimensional structures. The excellent features, such as high specificity and affinity for target proteins, small size, chemical stability, low immunogenicity, facile chemical synthesis, versatility in structural design and engineering, and accessible for site-specific modifications with functional moieties, make aptamers attractive molecules in the fields of clinical diagnostics and biopharmaceutical therapeutics. However, difficulties in purification and structural identification of aptamers remain a major impediment to their broad clinical application. In this mini-review, we present the recently attractive developments regarding the purification and identification of aptamers. We also discuss the advantages, limitations, and prospects for the major methods applied in purifying and identifying aptamers, which could facilitate the application of aptamers.

show abstract

Section: Separation and Purification Of Oligonucleotidesmentioning

confidence: 99%

Recent Methods for Purification and Structure Determination of Oligonucleotides

Zhang

Wang

et al. 2016

IJMS

View full text Add to dashboard Cite

show abstract

“…Three different chromatographic columns with different functional groups (octadecyl, octadecyl with embedded polar groups and pentauorophenyl) were selected for this study based on the literature, as well as on our previous experience in ASO analysis. 9,24,29,30 As it can be seen in Fig. 2, S2, S3 and Table 2, the highest k 0 values for ASOs were obtained for F5 column (Fig.…”

Section: Inuence Of Stationary Phases On Aso Retentionmentioning

confidence: 63%

Analysis of antisense oligonucleotides with the use of ionic liquids as mobile phase modifiers

et al. 2019

View full text Add to dashboard Cite

The main goal of this study was the investigation of the impact of several ionic liquids, commonly used as free silanol suppressors, on the retention and separation of phosphorothioate oligonucleotides. Three various stationary phases (octadecyl, octadecyl with embedded polar groups and pentafluorophenyl) as well as ionic liquids with the concentration range of 0.1-7 mM were used for this purpose. The results obtained during this study showed that the increase in concentration of ionic liquids results in increasing retention of the oligonucleotides. Such an effect was observed regardless of the stationary phase used.Moreover, elongation of the alkyl chain in the structure of ionic liquids caused an increase of antisense oligonucleotide retention factors. The results obtained during retention studies confirmed that addition of ionic liquids to the mobile phase influences antisense oligonucleotide retention in a way similar to the case of commonly used ion pair reagents such as amines. A method of oligonucleotide separation was also developed. The best selectivity was obtained for the octadecyl stationary phase since separation of mixtures of antisense oligonucleotides and their metabolites differing in sequence length was successful.It has to be pointed out that ionic liquids were used for the first time as mobile phase additives for oligonucleotide analysis. View Article Online AE 0.443 29.255 AE 0.370 36.562 AE 0.198 41.360 AE 0.468 41.623 AE 0.000 OL2 13.135 AE 0.075 15.210 AE 0.148 16.695 AE 0.223 n.a. n.a. 23.688 AE 0.000 25.679 AE 0.000 28.172 AE 0.025 35.761 AE 0.395 39.686 AE 0.171 40.000 AE 0.121 OL3 10.921 AE 0.097 13.207 AE 0.025 14.862 AE 0.000 n.a. n.a. 21.666 AE 0.050 23.933 AE 0.050 26.341 AE 0.000 34.697 AE 0.122 37.768 AE 0.025 39.426 AE 0.199 OL4 14.846 AE 1.532 15.892 AE 0.075 17.235 AE 0.050 n.a. n.a. 24.684 AE 0.025 26.743 AE 0.025 29.080 AE 0.025 37.401 AE 0.50 41.238 AE 0.050 42.336 AE 0.075 OL5 13.257 AE 0.050 15.490 AE 0.050 16.780 AE 0.050 n.a. n.a. 24.282 AE 0.050 26.357 AE 0.025 28.677 AE 0.000 37.542 AE 0.000 41.222 AE 0.025 42.983 AE 0.345 OL6 11.583 AE 0.148 13.973 AE 0.079 15.524 AE 0.000 n.a. n.a. 23.585 AE 0.050 23.986 AE 2.738 28.313 AE 0.025 37.768 AE 0.075 41.291 AE 0.025 43.263 AE 0.050 This journal isFig. 5 Chromatograms obtained for: (A) OL4 extracted from enriched serum sample, (B) separation of OL1 and its potential sequence related impurity. Chromatographic conditions: (A) Kinetex F5 stationary phase, gradient elution program: 45-60% v/v of MeOH in 10 minutes, (B) syncronis aQ stationary phase, gradient elution program: 48-55% v/v of MeOH in 10 minutes. UV detection at l ¼ 260 nm, autosampler and column temperature: 30 C, mobile phase flow rate 0.3 ml min À1 . Injection volume: 2 ml. 39108 | RSC Adv., 2019, 9, 39100-39110 This journal is

show abstract

“…In the vast majority of cases, C18 stationary phases are employed for the separation of ONs in IP-RPLC [74]. Recently, other adsorbents have also been considered.…”

Section: Analytical Characterization Of Oligonucleotidesmentioning

confidence: 99%

“…High performance Liquid Chromatography (LC) is indeed the technique of choice for this.Since ONs can bear an elevated number of negative charges depending on their length, the most effective elution modes are ion-pair reversed-phase liquid chromatography (IP-RPLC), anion exchange (AEX) LC, hydrophilic interaction liquid chromatography (HILIC) and mixed-mode LC[74]. Detection of ONs is usually performed through a UV detector, typically at 260 nm, where they exhibit very strong absorbance by their heterocyclic ring.…”

mentioning

confidence: 99%

See 1 more Smart Citation

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

Catani

Luca

Alcântara

et al. 2020

Biotechnology Journal

View full text Add to dashboard Cite

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.

show abstract

Liquid chromatographic separation of oligonucleotides

Cited by 9 publications

References 81 publications

Recent Methods for Purification and Structure Determination of Oligonucleotides

Recent Methods for Purification and Structure Determination of Oligonucleotides

Analysis of antisense oligonucleotides with the use of ionic liquids as mobile phase modifiers

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

Contact Info

Product

Resources

About