Quantitative target engagement of RIPK1 in human whole blood via the cellular thermal shift assay for potential pre-clinical and clinical applications.
SLAS Discov
; 29(2): 100135, 2024 Mar.
Article
en En
| MEDLINE
| ID: mdl-38101572
ABSTRACT
The cellular thermal shift assay (CETSA®) is a target engagement method widely used for preclinical characterization of small molecule compounds. CETSA® has been used for semi-quantitative readouts in whole blood with PBMC isolation, and quantitative, plate-based readouts using cell lines. However, there has been no quantitative evaluation of CETSA® in unprocessed human whole blood, which is preferred for clinical applications. Here we report two separate assay formats - Alpha CETSA® and MSD CETSA® - that require less than 100 µL of whole blood per sample without PBMC isolation. We chose RIPK1 as a proof-of-concept target and, by measuring engagement of seven different inhibitors, demonstrate high assay sensitivity and robustness. These quantitative CETSA® platforms enable possible applications in preclinical pharmacokinetic-pharmacodynamic studies, and direct target engagement with small molecules in clinical trials.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Bioensayo
/
Leucocitos Mononucleares
Límite:
Humans
Idioma:
En
Revista:
SLAS Discov
Año:
2024
Tipo del documento:
Article
País de afiliación:
Estados Unidos