| Title |
Degradation product characterization of therapeutic oligonucleotides using liquid chromatography mass spectrometry
|
|---|---|
| Published in |
Analytical & Bioanalytical Chemistry, April 2018
|
| DOI | 10.1007/s00216-018-1032-8 |
| Pubmed ID | |
| Authors |
N. M. Elzahar, N. Magdy, Amira M. El-Kosasy, Michael G. Bartlett |
| Abstract |
Synthetic antisense phosphorothioate oligonucleotides (PS) have undergone rapid development as novel therapeutic agents. The increasing significance of this class of drugs requires significant investment in the development of quality control methods. The determination of the many degradation pathways of such complex molecules presents a significant challenge. However, an understanding of the potential impurities that may arise is necessary to continue to advance these powerful new therapeutics. In this study, four different antisense oligonucleotides representing several generations of oligonucleotide therapeutic agents were evaluated under various stress conditions (pH, thermal, and oxidative stress) using ion-pairing reversed-phase liquid chromatography tandem mass spectrometry (IP-RPLC-MS/MS) to provide in-depth characterization and identification of the degradation products. The oligonucleotide samples were stressed under different pH values at 45 and 90 °C. The main degradation products were observed to be losses of nucleotide moieties from the 3'- and 5'-terminus, depurination, formation of terminal phosphorothioates, and production of ribose, ribophosphorothioates (Rp), and phosphoribophosphorothioates (pRp). Moreover, the effects of different concentrations of hydrogen peroxide were studied resulting in primarily extensive desulfurization and subsequent oxidation of the phosphorothioate linkage to produce the corresponding phosphodiester. The reaction kinetics for the degradation of the oligonucleotides under the different stress conditions were studied and were found to follow pseudo-first-order kinetics. Differences in rates exist even for oligonucleotides of similar length but consisting of different sequences. Graphical abstract Identification of degradation products across several generations of oligonucleotide therapeutics using LC-MS. |
X Demographics
The data shown below were collected from the profiles of 4 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 2 | 50% |
| Unknown | 2 | 50% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 2 | 50% |
| Science communicators (journalists, bloggers, editors) | 1 | 25% |
| Members of the public | 1 | 25% |
Mendeley readers
The data shown below were compiled from readership statistics for 38 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 38 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 5 | 13% |
| Researcher | 4 | 11% |
| Student > Master | 3 | 8% |
| Student > Doctoral Student | 2 | 5% |
| Student > Bachelor | 2 | 5% |
| Other | 4 | 11% |
| Unknown | 18 | 47% |
| Readers by discipline | Count | As % |
|---|---|---|
| Pharmacology, Toxicology and Pharmaceutical Science | 8 | 21% |
| Chemistry | 6 | 16% |
| Agricultural and Biological Sciences | 3 | 8% |
| Biochemistry, Genetics and Molecular Biology | 1 | 3% |
| Unknown | 20 | 53% |
Attention Score in Context
This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 11 May 2018.
All research outputs
#16,728,456
of 25,382,440 outputs
Outputs from Analytical & Bioanalytical Chemistry
#5,260
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Outputs of similar age
#209,447
of 341,757 outputs
Outputs of similar age from Analytical & Bioanalytical Chemistry
#75
of 177 outputs
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We're also able to compare this research output to 177 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 53% of its contemporaries.