MALDI-TOF Mass Spectrometry-Based Assay for Measuring Covalent Target Engagement of KRAS G12C Inhibitors

Marcin Dyba; John-Paul Denson; Anna E Maciag

doi:10.1007/978-1-0716-3822-4_11

MALDI-TOF Mass Spectrometry-Based Assay for Measuring Covalent Target Engagement of KRAS G12C Inhibitors

Methods Mol Biol. 2024:2797:145-157. doi: 10.1007/978-1-0716-3822-4_11.

Authors

Marcin Dyba¹, John-Paul Denson², Anna E Maciag²

Affiliations

¹ NCI RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, USA. marcin.dyba@nih.gov.
² NCI RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.

PMID: 38570458
DOI: 10.1007/978-1-0716-3822-4_11

Abstract

MALDI-TOF mass spectrometry enables high-throughput screening of covalent fragment libraries and SAR compound progressions of selective KRAS G12C inhibitors. Using the MALDI-TOF platform instead of the more traditional ESI-MS TOF/orbitrap instrumentation can radically shorten sample acquisition time, allowing up to 384 samples to be screened in 30 min. The typical throughput for a covalent library screen is 1152 samples per 8 h, including processing, calculation, and reporting steps. The throughput can be doubled without any significant assay modification.

Keywords: Adagrasib; Covalent screening; Fragment-based screening; High-throughput screening; KRAS G12C; MALDI-TOF MS; RAS covalent inhibitors; Sotorasib.

MeSH terms

High-Throughput Screening Assays* / methods
Mutation
Proto-Oncogene Proteins p21(ras)* / genetics
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods

Substances

Proto-Oncogene Proteins p21(ras)