Quantitative target engagement of RIPK1 in human whole blood via the cellular thermal shift assay for potential pre-clinical and clinical applications.

Autor:	Patel S; AbbVie Inc., North Chicago, IL 60064, USA., Karlsson M; Pelago Bioscience AB, Solna, Sweden., Klahn JT; AbbVie Inc., North Chicago, IL 60064, USA., Gambino F Jr; AbbVie Inc., North Chicago, IL 60064, USA., Costa H; Pelago Bioscience AB, Solna, Sweden., McGuire KA; AbbVie Inc., North Chicago, IL 60064, USA., Baumgartner CK; AbbVie Inc., North Chicago, IL 60064, USA., Williams J; AbbVie Inc., North Chicago, IL 60064, USA., Sandoz S; Pelago Bioscience AB, Solna, Sweden. Electronic address: sarah.sandoz@pelagobio.com., Kath JE; AbbVie Inc., North Chicago, IL 60064, USA. Electronic address: james.kath@abbvie.com.
Jazyk:	angličtina
Zdroj:	SLAS discovery : advancing life sciences R & D [SLAS Discov] 2024 Mar; Vol. 29 (2), pp. 100135. Date of Electronic Publication: 2023 Dec 14.
DOI:	10.1016/j.slasd.2023.12.007
Abstrakt:	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. Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: H.C and S.S. are employees of Pelago Bioscience AB. M.K. is an author who was an employee of Pelago, passed away prior to publication, and is listed with the agreement of all other authors. S.P., J.T.K, F.G., K.A.M., C.K.B., J.W., and J.E.K. are employees of AbbVie Inc. J.T.K, K.A.M., C.K.B., and J.E.K. hold stock in AbbVie Inc. Pelago controls the global patent portfolio protecting the CETSA® technology. CETSA® data was generated at AbbVie under license from Pelago. The design, study conduct, and financial support for this research were provided by AbbVie and Pelago. AbbVie and Pelago participated in the interpretation of data, review, and approval of the publication. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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