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Using High Content Imaging to Quantify Target Engagement in Adherent Cells

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Identifying drug targets with thermal proteome profiling using IBT-16plex.

Zhaomei Shi¹, Yan Ren², Shuwei Li^3,4

¹School of Life Science and Technology, ShanghaiTech University, Shanghai, China.

Rapid Communications in Mass Spectrometry : RCM

|December 11, 2023

Summary

A new reagent, IBT-16plex, effectively identifies drug targets using thermal proteome profiling (TPP). This cost-effective method offers improved protein identification and quantification compared to TMTpro-16plex.

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Area of Science:

Chemical Biology
Proteomics
Drug Discovery

Background:

Thermal proteome profiling (TPP) is a key technique for identifying drug targets.
TMTpro-16plex facilitates TPP by enabling single-batch processing of multiple samples.
A novel isobaric labeling reagent, IBT-16plex, was developed to enhance TPP performance.

Purpose of the Study:

To evaluate the efficacy of the novel IBT-16plex reagent for drug target identification using TPP.
To compare the performance of IBT-16plex against the established TMTpro-16plex reagent.

Main Methods:

Application of IBT-16plex for thermal proteome profiling (TPP).
Target identification for methotrexate and panobinostat.
Analysis of protein thermal stability shifts to identify drug-target interactions.

Main Results:

Successful identification of known drug targets for methotrexate and panobinostat.
Detection of elevated melting temperatures for identified targets.
Identification of known and potential novel drug off-targets.

Conclusions:

IBT-16plex demonstrates superior or comparable performance to TMTpro-16plex in TPP.
IBT-16plex is a viable and cost-effective alternative to TMTpro-16plex for TPP applications.
The study validates IBT-16plex for robust drug target and off-target identification.