Informace o publikaci
Tools that improve concentration sensitivity in capillary electrophoresis-frontal analysis for affinity studies
| Autoři | |
|---|---|
| Rok publikování | 2024 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | Affinity interaction studies are important for proper understanding biomolecular interaction mechanism. Pharmaceutical industry during new drug development in lead optimization considers plasma protein–drug interaction and its influence on drug distribution in the patient organism. Plasma protein–drug (small molecule) interaction is low binding interactions with fast kinetics. Due to this fact complex could not be detected as entity. The most abundant protein is human serum albumin (HSA). The interaction of drugs with HSA strongly influences free drug concentration which enables therapeutic effect. Capillary electrophoresis-frontal analysis (CE-FA) is suitable method for studying this type of interaction. Measurement of binding parameters does not necessarily require pure samples as well as immobilisation neither derivatisation of binding partners. The method save the biological material, samples and reagents with its miniaturisation. However, the biggest challenge of CE-FA method is its low concentration sensitivity linked with commercially incorporated UV-VIS detector and narrow capillary diameter around 10 – 100 µm. This study offers newly optimised methods with enhanced concentration sensitivity of CE-FA method for HSA and small molecule drugs interaction studies. Newly developed methods are CE-FA UV-VIS with on-line preconcentration technique (field amplified sample stacking) and CE-FA combined with more sensitive detectors as contactless conductivity or MS detector. The obtained binding parameters binding constant and number of binding sites are compared and corresponds with values range from optimised conventional CE-FA UV-VIS method and different studies from literature. Newly optimized method contributes with higher sensitivity, universality and extents CE usage. |