| Autor: |
Daniel Muthas, John Simpson, Linda Yrlid, Samantha Peel, Claus Bendtsen, Keith R. Willison, Suhuur Osman, David R. Klug |
| Přispěvatelé: |
Engineering & Physical Science Research Council (EPSRC) |
| Rok vydání: |
2021 |
| Předmět: |
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| Popis: |
The cellular thermal shift assay (CETSA) has been used extensively since its introduction to study drug-target engagement within both live cells and cellular lysate. This has proven to be a useful tool in early stage drug discovery and is used to study a wide range of protein classes. We describe the application of a single-cell CETSA workflow within a microfluidic affinity capture (MAC) chip. This has enabled us to quantitatively determine the active FOXO1 single-molecule count and observe FOXO1 stabilization and destabilization in the presence of three small molecule inhibitors, including demonstrating the determination of EC50. The successful use of the MAC chip for single-cell CETSA paves the way for the study of precious clinical samples owing to the low number of cells needed by the chip. It also provides a useful tool for studying any underlying population heterogeneity that exists within a cellular system, a feature that is usually masked when conducting ensemble measurements. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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