Synthesis and application of stationary phase for DNA-affinity chromatographic analysis of unmodified and antisense oligonucleotide
| Autor: | Ewelina Zawadzka, Sylwia Studzińska, Michał Szumski, Szymon Bocian |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Ion chromatography
Stacking Oligonucleotides Liquid chromatography 02 engineering and technology 010402 general chemistry 01 natural sciences Biochemistry Chromatography Affinity DNA Antisense Analytical Chemistry chemistry.chemical_compound Affinity chromatography Selectivity Chromatography Chemistry Oligonucleotide Hydrophilic interaction chromatography Correction DNA DNA-affinity chromatography Chromatography Ion Exchange 021001 nanoscience & nanotechnology 0104 chemical sciences Retention Polynucleotide Oligonucleotide-based stationary phase 0210 nano-technology Research Paper |
| Zdroj: | Analytical and Bioanalytical Chemistry Anal Bioanal Chem |
| ISSN: | 1618-2650 1618-2642 |
| Popis: | Graphical abstract The goal of the research was the synthesis and application of an oligonucleotide immobilized stationary phase for the analysis of unmodified and antisense oligonucleotides. The method for attaching these molecules to aminopropyl silica modified with pentanedioic acid was developed. Each step of the synthesis was carefully controlled with the application of spectroscopic, elemental, and chromatographic analyses. The oligonucleotide-based stationary phase was applied for the retention studies. Unmodified oligonucleotides of different complementarity to the molecule attached as a stationary phase, as well as antisense oligonucleotides, were tested. The comparative study upon complex optimization of oligonucleotide analysis in different liquid chromatography modes was performed. Results have shown that this stationary phase may be applied for oligonucleotide analysis in hydrophilic interaction liquid chromatography and ion exchange chromatography, but no unique sequence-based selectivity was obtained. Contrary results were observed for affinity chromatography, which allowed for specific separation of the complementary strands based on hydrogen bonding and stacking interactions, where the temperature was the main factor influencing the selectivity of the separation. Furthermore, the oligonucleotide-based stationary phase may be applied for comparative antisense oligonucleotide hybridization studies to a specific RNA sequence. All of the results have shown that affinity chromatography with oligonucleotide-based stationary phases is a powerful technique for the specific base recognition of polynucleotides. Supplementary Information The online version contains supplementary material available at 10.1007/s00216-021-03473-7. |
| Databáze: | OpenAIRE |
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