Experimental strategies to improve drug-target identification in mass spectrometry-based thermal stability assays

ABSTRACT: In this study, we developed a model system for the assessment of small molecule-protein interactions using intact cells treated with a commercially available highly specific mitogen-activated protein kinase (MEK) 1/2 inhibitor to prepare a set of unfractionated test samples labeled with tandem mass tags. Our objective was to improve qualitative and quantitative aspects of MS-TSA as well as its efficiency and accuracy. We evaluated individually and for the first time in combination ΦSDM, FAIMS, and SIILCC as an improved MS-based acquisition approach for thermal stability assays (iMAATSA). PSM-level filtering was preliminarily investigated as an approach to reduce melting curve variation and improve the accuracy of Tm measurements.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): T Cell, Cell Culture

SUBMITTER: Clifford Phaneuf  

LAB HEAD: Prof. Alexander Ivanov

PROVIDER: PXD034087 | Pride | 2023-05-10

REPOSITORIES: Pride