Salt gradient and ion-pair mediated anion exchange of intact messenger ribonucleic acids
Small molecule drugs and protein-based therapeutics are widely used to treat various diseases, yet even more aspects of human health can be addressed through new modalities, like messenger RNA (mRNA). The complex nature and chemical structure of an mRNA molecule will require the development of new a...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-11-01
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| Series: | Journal of Chromatography Open |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2772391722000020 |
| _version_ | 1828092081791303680 |
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| author | Szabolcs Fekete Hua Yang Kevin Wyndham Matthew Lauber |
| author_facet | Szabolcs Fekete Hua Yang Kevin Wyndham Matthew Lauber |
| author_sort | Szabolcs Fekete |
| collection | DOAJ |
| description | Small molecule drugs and protein-based therapeutics are widely used to treat various diseases, yet even more aspects of human health can be addressed through new modalities, like messenger RNA (mRNA). The complex nature and chemical structure of an mRNA molecule will require the development of new analytical methods. In this work, we propose the use of ion exchange and relatively mild elution conditions as the basis for alternative analytical methods. We have found that a gradient of weak ion-pairing cations can produce intriguing AEX separations of mRNA. This so-called ion pairing anion exchange “IPAX” method provides different recovery and selectivity effects compared to classical sodium chloride salt gradients. We have also shown there to be value in combining the use of an ion-pairing agent as a mobile phase additive along with the implementation of a classical salt gradient. It might be most appropriate to refer to this separation technique as an “ion-pairing mediated salt gradient”. The column temperature has been found to be a very important factor that should be applied to test for effects on various chromatographic values and to learn about an mRNA molecule while confirming its optimal separation conditions. In a classical salt gradient separation, relatively low temperatures (T = 30–40 °C) resulted in better recoveries while in the IPAX separation, higher temperatures up to 60 °C seemed to be more beneficial. An ion-pairing mediated salt gradient appeared to be best matched to temperatures between 30 and 45 °C. Plotting van't Hoff data has also elucidated new information and that a classical salt gradient seems to be an entropy driven process, while the IPAX separation seems to be more complex. It exhibits an entropy driven process at lower temperatures and then an enthalpy controlled one at higher temperatures. |
| first_indexed | 2024-04-11T06:22:42Z |
| format | Article |
| id | doaj.art-3405dd961f9647a9becbd051f5cf7fe0 |
| institution | Directory Open Access Journal |
| issn | 2772-3917 |
| language | English |
| last_indexed | 2024-04-11T06:22:42Z |
| publishDate | 2022-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Chromatography Open |
| spelling | doaj.art-3405dd961f9647a9becbd051f5cf7fe02022-12-22T04:40:29ZengElsevierJournal of Chromatography Open2772-39172022-11-012100031Salt gradient and ion-pair mediated anion exchange of intact messenger ribonucleic acidsSzabolcs Fekete0Hua Yang1Kevin Wyndham2Matthew Lauber3Waters Corporation, located in CMU-Rue Michel Servet 1, 4, Geneva 1211, Switzerland; Corresponding author.Waters Corporation, 34 Maple Street, Milford, MA 01757, United StatesWaters Corporation, 34 Maple Street, Milford, MA 01757, United StatesWaters Corporation, 34 Maple Street, Milford, MA 01757, United StatesSmall molecule drugs and protein-based therapeutics are widely used to treat various diseases, yet even more aspects of human health can be addressed through new modalities, like messenger RNA (mRNA). The complex nature and chemical structure of an mRNA molecule will require the development of new analytical methods. In this work, we propose the use of ion exchange and relatively mild elution conditions as the basis for alternative analytical methods. We have found that a gradient of weak ion-pairing cations can produce intriguing AEX separations of mRNA. This so-called ion pairing anion exchange “IPAX” method provides different recovery and selectivity effects compared to classical sodium chloride salt gradients. We have also shown there to be value in combining the use of an ion-pairing agent as a mobile phase additive along with the implementation of a classical salt gradient. It might be most appropriate to refer to this separation technique as an “ion-pairing mediated salt gradient”. The column temperature has been found to be a very important factor that should be applied to test for effects on various chromatographic values and to learn about an mRNA molecule while confirming its optimal separation conditions. In a classical salt gradient separation, relatively low temperatures (T = 30–40 °C) resulted in better recoveries while in the IPAX separation, higher temperatures up to 60 °C seemed to be more beneficial. An ion-pairing mediated salt gradient appeared to be best matched to temperatures between 30 and 45 °C. Plotting van't Hoff data has also elucidated new information and that a classical salt gradient seems to be an entropy driven process, while the IPAX separation seems to be more complex. It exhibits an entropy driven process at lower temperatures and then an enthalpy controlled one at higher temperatures.http://www.sciencedirect.com/science/article/pii/S2772391722000020Ion exchange chromatographyIon-pairMessenger ribonucleic acidmRNAVaccinesGene therapy |
| spellingShingle | Szabolcs Fekete Hua Yang Kevin Wyndham Matthew Lauber Salt gradient and ion-pair mediated anion exchange of intact messenger ribonucleic acids Journal of Chromatography Open Ion exchange chromatography Ion-pair Messenger ribonucleic acid mRNA Vaccines Gene therapy |
| title | Salt gradient and ion-pair mediated anion exchange of intact messenger ribonucleic acids |
| title_full | Salt gradient and ion-pair mediated anion exchange of intact messenger ribonucleic acids |
| title_fullStr | Salt gradient and ion-pair mediated anion exchange of intact messenger ribonucleic acids |
| title_full_unstemmed | Salt gradient and ion-pair mediated anion exchange of intact messenger ribonucleic acids |
| title_short | Salt gradient and ion-pair mediated anion exchange of intact messenger ribonucleic acids |
| title_sort | salt gradient and ion pair mediated anion exchange of intact messenger ribonucleic acids |
| topic | Ion exchange chromatography Ion-pair Messenger ribonucleic acid mRNA Vaccines Gene therapy |
| url | http://www.sciencedirect.com/science/article/pii/S2772391722000020 |
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