Competitive Activity-Based Protein Profiling (ABPP) Service
MtoZ Biolabs provides a Competitive Activity-Based Protein Profiling (ABPP) Service to evaluate small molecule target engagement and selectivity across enzyme families using activity-based probes and LC-MS/MS readouts. The service supports mechanism-of-action studies, lead optimization, and off-target risk assessment for covalent and competitive inhibitors by translating compound binding into proteome-resolved activity changes.
What Is Competitive Activity-Based Protein Profiling (ABPP)?
Competitive Activity-Based Protein Profiling is a chemoproteomics approach used to measure small molecule target engagement directly in complex biological samples such as cells, tissues, or lysates. Rather than inferring binding from phenotype or protein abundance, competitive ABPP reads out functional occupancy at an enzyme active site or activity-linked residue, which makes it well suited for enzyme families where closely related proteins are hard to distinguish by conventional assays.
Competitive ABPP is widely used to answer practical questions in discovery and chemical biology. It helps confirm whether a compound engages the intended targets in cells, reveals selectivity across related enzymes, and flags potential off-target liabilities early. Because the readout is activity-centered, it can capture regulation that is missed by total proteome measurements, providing decision-ready evidence for mechanism-of-action studies and lead optimization.
Principle of Competitive Activity-Based Protein Profiling (ABPP)
The principle is straightforward. A sample is first incubated with your test compound, allowing it to bind to its targets under near-native conditions. An activity-based probe is then added to covalently label active enzymes within the probe-covered class. If the compound occupies the same functional site, probe labeling decreases in a dose-dependent manner compared with a vehicle control. This loss of labeling serves as a direct signature of target engagement, and the competed proteins can be visualized by gel-based readouts or identified and quantified at scale by LC-MS/MS.
Zhu, H. et al. Isr J Chem. 2023.
Figure 1. Principle of competitive ABPP
Competitive Activity-Based Protein Profiling (ABPP) Service at MtoZ Biolabs
MtoZ Biolabs offers a flexible, project-driven service that can be configured for discovery, selectivity profiling, or focused validation. Probe chemistry and readout type are selected based on your compound class, target hypothesis, and biological matrix.
Our service include:
💠Gel-based competitive ABPP for rapid screening and visual engagement confirmation.
💠MS-based competitive ABPP for identification and quantification of competed targets at scale.
💠Targeted follow-up options for higher precision on a defined set of enzymes.
Workflow of Competitive Activity-Based Protein Profiling (ABPP) Service
Zhu, H. et al. Isr J Chem. 2023.
Figure 2. General Workflow for LC-MS/MS-Based Competitive ABPP
Why Choose MtoZ Biolabs
✔️Advanced Platforms
MtoZ Biolabs established an advanced Competitive Activity-Based Protein Profiling (ABPP) Service platform, guaranteeing reliable, fast, and highly accurate analysis service.
✔️Deep Technical Expertise
Experienced scientists guide probe strategy selection, control design, and interpretation to avoid common pitfalls.
✔️Customizable Solutions
Project design can be tuned for discovery screening, selectivity profiling, or focused target validation.
✔️High-Quality Data
QC checkpoints, appropriate blanks, and reproducible workflows support confidence in comparisons across groups.
✔️Outstanding Customer Service
Responsive communication and clear reporting help teams move quickly from results to next-step decisions.
Applications of Competitive Activity-Based Protein Profiling (ABPP) Service
1. Drug Discovery and Lead Optimization
Compound engagement and selectivity can be measured directly in cells or tissues, supporting SAR-driven compound ranking and lead refinement.
2. Mechanism-of-Action and Target Deconvolution
Phenotypic activity can be traced back to functional target occupancy by identifying which probe-covered enzymes are competed under treatment conditions.
3. Off-Target Risk Assessment
Proteome-resolved competition profiles help uncover unintended enzyme engagement that may drive liabilities or confound interpretation.
4. Translational and pharmacodynamic research
Engagement signatures provide practical evidence to connect exposure with target occupancy and to guide biomarker selection for follow-up studies.
FAQ
Q1: What types of samples are suitable?
1. Cultured cells or primary cells from defined conditions
2. Fresh or frozen tissues
3. Biofluids such as plasma, serum, CSF, or other biological fluids when applicable
4. Cell lysates
5. Purified proteins or enzyme preparations for focused target engagement studies
Q2: How should I prepare my samples?
1. Keep collection and handling consistent across groups and work cold when possible to preserve activity
2. Avoid harsh denaturants and unnecessary detergents, and share buffer composition and any inhibitors used
3. Aliquot samples to prevent repeated freeze-thaw cycles, label clearly, store at -80°C, and ship on dry ice
4. Provide a sample sheet with sample IDs, conditions, replicates, and your research objective for study alignment
For more information, please refer to Sample Submission Guidelines for Proteomics and Sample Submission Guidelines for Metabolomics.
Q3: What is the service general workflow?
Q4: What data formats are provided?
1. Raw data: vendor MS raw files plus optional mzML; gel or blot images when gel-based readouts are included
2. Processed tables: XLSX or CSV (protein IDs with gene symbols, peptide evidence, quantitative metrics, enrichment versus controls, competition effect sizes, and statistics when applicable)
3. Figures and QC: PDF plus PNG or TIFF (QC summaries, replicate concordance, enrichment and competition plots, heatmaps, and network views when applicable)
4. Report and methods: concise PDF describing assay design, probe and competitor conditions, key parameters, results summary, and interpretation notes, plus instrument settings and software versions for reproducibility
Additional formats can be provided upon request to meet specific analysis or publication requirements.
Start Your Project with MtoZ Biolabs
MtoZ Biolabs, an integrated Chromatography and Mass Spectrometry (MS) Services Provider, provides advanced proteomics, metabolomics, and biopharmaceutical analysis services to researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our Competitive Activity-Based Protein Profiling (ABPP) Service is designed to provide more rapid, high-throughput, and cost-effective analysis, with exceptional data quality and minimal sample consumption. Contact us to discuss your experimental design or request a quote.
Related Services
How to order?
