Our company has established a label-free target engagement analysis platform based on thermal stability profiling approaches for bioactive natural products and small molecules. By monitoring ligand-induced changes in protein thermal stability under physiologically relevant conditions, this platform supports target identification, target validation, and mechanism-of-action studies in complex biological systems. The platform is suitable for intact cells, cell lysates, tissue homogenates, and related biological samples, enabling researchers to evaluate compound–protein interactions without chemical labeling or immobilization.
Principle
Protein–ligand interactions frequently alter the conformational stability of proteins. Upon heating, proteins progressively unfold and aggregate, resulting in reduced solubility. When a compound interacts with a target protein, the thermal stability of that protein may change, which can be detected by comparing soluble protein abundance across temperature conditions. By combining controlled thermal perturbation with downstream protein quantification methods, thermal stability profiling enables the assessment of compound–target engagement in biologically relevant environments. This approach can be applied to purified proteins, cell lysates, intact cells, tissues, and in vivo–derived samples, providing valuable support for target discovery and pharmacological studies.
Our Technology Platform
Our thermal stability profiling workflow integrates multiple analytical strategies for compound–target interaction studies, including:
- Label-free target engagement analysis
- Medium- and high-throughput thermal stability workflows
- Western blot–based protein detection
- Immunoassay-based protein quantification
- Mass spectrometry–assisted protein profiling
- Proteome-scale thermal response analysis
By integrating thermal stability measurements with quantitative proteomics approaches, our platform supports both hypothesis-driven target validation and broader exploratory target discovery studies.
Typical Workflow
Step 1
Experimental Strategy
Based on the biological question and physicochemical properties of the compounds, appropriate experimental formats, biological models, temperature conditions, and detection approaches are selected.
Step 2
Sample Preparation
Cell lysates, tissue homogenates, or intact-cell systems are prepared under conditions designed to preserve endogenous protein structures and interactions.
Step 3
Compound Incubation
Bioactive compounds or natural product samples are incubated with biological materials to allow potential compound–protein interactions to occur.
Step 4
Thermal Treatment
Samples are exposed to controlled temperature conditions to induce protein unfolding and aggregation. Differences in protein thermal behavior between treated and control groups are subsequently evaluated.
Step 5
Separation of Soluble Fractions
Following thermal treatment, insoluble aggregated proteins are removed by centrifugation, and soluble protein fractions are collected for downstream analysis.
Step 6
Protein Detection and Quantification
Protein abundance in soluble fractions can be analyzed using western blotting, immunodetection methods, or mass spectrometry–based workflows, depending on project objectives.
Step 7
Data Analysis
Thermal response curves are generated by comparing protein abundance across temperature conditions. Statistically significant differences may indicate compound-associated thermal stability changes.
Expanded Applications of Platform
Our platform not only supports target identification, but also enables a range of downstream studies, including:
- Mechanism-of-action (MoA) studies
- Target deconvolution for phenotypic screening hits
- Lead optimization and structure–activity relationship (SAR) studies
- Confirmation of on-target activity in relevant disease models
By leveraging this technology, our company provides a powerful, label-free solution for target identification and validation of bioactive natural products. Our integrated platform delivers physiologically relevant insights, enabling researchers to confidently advance compounds from discovery to development.
Online Inquiry
Frequently Asked Questions (FAQ)
Q1: Can this approach be used for weak or transient interactions?
Q2: What sample types are supported?
A: We support cell lines, primary cells, tissues, and in vivo samples, depending on project requirements.
Q3: Can the data be integrated with other omics approaches?
A: Yes. Thermal stability profiling data may be combined with transcriptomics, metabolomics, proteomics, and functional assays for broader biological interpretation.
Disclaimer
This webpage describes independent thermal stability–based analytical workflows for research applications. References to scientific concepts or commonly used assay methodologies are for descriptive and informational purposes only. Our company is not affiliated with, endorsed by, or sponsored by any third-party trademark holder. Any trademarks referenced remain the property of their respective owners.