Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Scalable hypothalamic neuron differentiation from human pluripotent stem cells suitable for modeling metabolic disorders.

Stem cell reports·2026
Same author

Integrating AI in Medicinal Chemistry for Accelerated Drug Discovery: A Comprehensive SAR (CSAR) Optimization Strategy and Discovery of Potent ALDH3A1 Inhibitors.

Journal of medicinal chemistry·2026
Same author

Co-Targeting Nuclear Export and Translation Initiation Uncovers a Therapeutic Vulnerability in Lethal Prostate Cancer.

bioRxiv : the preprint server for biology·2026
Same author

Correction to "Perfluorinated Iridium Catalyst for Signal Amplification by Reversible Exchange Provides Metal-Free Aqueous Hyperpolarized [1-<sup>13</sup>C]-Pyruvate".

Journal of the American Chemical Society·2026
Same author

Impact of Chemical Quality on High-Throughput in Vitro Assays: A Tox21 Study.

Chemical research in toxicology·2026
Same author

Ubiquitin-dependent recruitment of SLFN11 to chromatin is regulated by deubiquitinase (DUB) and RNF168.

bioRxiv : the preprint server for biology·2026

Related Experiment Video

Updated: Feb 18, 2026

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
13:22

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays

Published on: October 23, 2019

8.3K

Data-Independent Acquisition Enhancement of a Competitive Activity-Based Protein Profiling Platform for Kinase

Ravi Tharakan1, Yanyan Qu1, Michele Ceribelli1

  • 1National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland, USA.

Journal of Mass Spectrometry : JMS
|February 17, 2026
PubMed
Summary

This study introduces a new activity-based protein profiling (ABPP) platform using data-independent acquisition (DIA) mass spectrometry for efficient kinase drug target identification. The DIA approach significantly increases identified peptides and coverage while halving analysis time, enhancing kinase profiling workflows.

Keywords:
activity‐based protein profilingdata‐independent acquisitionkinasemass spectrometryproteomics

More Related Videos

Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

8.6K
Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
11:23

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

6.7K

Related Experiment Videos

Last Updated: Feb 18, 2026

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
13:22

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays

Published on: October 23, 2019

8.3K
Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

8.6K
Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
11:23

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

6.7K

Area of Science:

  • Biochemistry and Molecular Biology
  • Proteomics
  • Drug Discovery

Background:

  • Kinase inhibitors are crucial therapeutics in oncology and immunology.
  • Mass spectrometry (MS) with kinase-binding probes is key for identifying drug targets.
  • Data-independent acquisition (DIA) offers superior quantification over traditional MS methods like data-dependent acquisition (DDA).

Purpose of the Study:

  • To develop and validate an activity-based protein profiling (ABPP) platform utilizing DIA for kinase profiling.
  • To enhance the efficiency and scalability of kinase target identification workflows.
  • To compare the performance of DIA versus DDA for kinase profiling.

Main Methods:

  • Systematic development of an ABPP platform integrated with in-house informatics tools.
  • Application of DIA mass spectrometry for comprehensive peptide identification and quantification.
  • Evaluation of shorter liquid chromatography (LC) gradient times to assess throughput.

Main Results:

  • DIA resulted in over 100% increase in identified biotinylated peptides and >40% improved kinase peptide coverage compared to DDA.
  • Analysis time was reduced by 50% (90 min vs. 180 min per sample).
  • Shorter LC gradients did not significantly impact kinase peptide coverage, demonstrating DIA's scalability.

Conclusions:

  • The DIA-based ABPP platform provides a more effective and efficient workflow for kinase profiling.
  • This approach significantly enhances throughput and scalability for identifying kinase drug targets.
  • DIA mass spectrometry represents a powerful tool for advancing kinase inhibitor research and drug discovery.