For bioactive natural products, traditional target identification strategies often rely on chemical modification, labeling, or immobilization of compounds, which can alter their native activity, reduce binding affinity, or introduce experimental bias. To overcome these limitations, our company has established a bioactive natural products target identification technology platform centered on label-free biophysical methods. These approaches directly measure changes in protein stability, conformation, or thermodynamics upon ligand binding, enabling reliable identification of drug–target interactions under near-physiological conditions. By integrating multiple complementary biophysical technologies, we provide a robust and systematic solution for uncovering both direct and indirect protein targets of bioactive natural products.
What are Biophysical Methods?
Biophysical methods are label-free technologies for identifying drug targets by detecting changes in protein properties caused by ligand binding. These methods rely on the principle that ligands can influence protein stability, conformation, or dynamics by modifying their intramolecular non-covalent interactions. By comparing protein behavior before and after treatment with compounds, researchers can observe alterations such as increased proteolysis resistance, changes in thermal stability, shifts in unfolding equilibria, or variations in chemical reactivity, which indicate potential drug targets.
Unlike traditional affinity-based or chemical labeling approaches, biophysical methods do not require modification or immobilization of natural products, preserving their native structure and biological activity. These methods can be applied to purified proteins, cell lysates, and intact cells, enabling the identification of both direct-binding targets and indirectly affected proteins. As a result, biophysical methods provide a powerful, unbiased strategy for elucidating the mechanisms of action of bioactive natural products in drug discovery.
Explore Our Biophysical Methods Technology Platform
Our platform integrates several state-of-the-art biophysical technologies, each addressing different aspects of protein–ligand interactions. Together, they offer comprehensive coverage from proteome-wide target discovery to quantitative binding validation.
Drug Affinity Responsive Target Stability (DARTS) Technology
DARTS is based on the principle that ligand binding often protects target proteins from proteolytic degradation. Upon interaction with a natural product, the target protein adopts a more stable conformation, making it less susceptible to limited protease digestion. DARTS is particularly well suited for discovering direct-binding targets of natural products in complex biological samples.
Stability of Proteins from Rates of Oxidation (SPROX) Technology
SPROX measures ligand-induced changes in protein thermodynamic stability by monitoring the oxidation rates of methionine residues under chemical denaturation conditions. Binding of a natural product shifts the protein's unfolding equilibrium, which can be quantitatively detected. It is especially powerful for studying targets that may not show large thermal shifts but exhibit subtle stability changes.
Thermal Stability-Based Target Engagement Technology
It evaluates protein stability changes upon ligand binding by heating intact cells or lysates across a temperature gradient. Target engagement increases the thermal stability of the protein, leading to higher resistance to heat-induced aggregation. It allows target validation under physiologically relevant conditions, confirming whether a natural product truly engages its target inside cells rather than only in vitro.
Thermal Proteome Profiling (TPP) Technology
By combining thermal denaturation, quantitative mass spectrometry, and bioinformatics analysis, TPP enables the identification of hundreds to thousands of proteins whose thermal stability is altered upon compound treatment. TPP provides a global view of drug–protein interactions, revealing primary targets, off-targets, and pathway-level effects, making it invaluable for mechanism-of-action studies and safety profiling.
Isothermal Titration Calorimetry (ITC) Technology
ITC is a gold-standard biophysical technique for directly measuring the thermodynamics of molecular interactions. It quantifies binding affinity (Kd), stoichiometry, enthalpy (ΔH), and entropy (ΔS) without any labeling or immobilization. ITC is used for definitive validation of direct binding between natural products and their protein targets, supporting hit confirmation and lead optimization.
Instruments and Equipment at Our Platform
To ensure high data quality, reproducibility, and scalability of our biophysical target identification services, our platform is equipped with a comprehensive suite of advanced instruments and supporting infrastructure. Each piece of equipment is carefully selected to support label-free biophysical analyses from proteome-wide discovery to high-precision target validation.
- High-resolution mass spectrometry systems
- Nano-LC and UHPLC systems
- Automated thermal control and heating systems
- Isothermal titration calorimeters (ITC)
- High-speed and ultracentrifuges
- Automated liquid handling and sample preparation systems
- Protein expression and purification systems
- Cell culture facilities
- Spectrophotometers and plate readers
- Advanced data analysis and bioinformatics infrastructure
Our Advantages
- Label-Free and Native Conditions: No chemical modification of natural products is required.
- Integrated Multi-Technology Platform: Complementary methods reduce false positives and increase confidence.
- Proteome-Wide and Target-Specific Capabilities: From discovery to validation in one platform.
- Expert Scientific Team: Deep expertise in natural products, biophysics, and mass spectrometry.
- Customizable Solutions: Tailored workflows based on compound properties and research goals.
The advent of biophysical methods for target identification has compensated for the deficiencies of chemical probe approach. Through our comprehensive biophysical methods technology platform, we empower researchers to unlock the full therapeutic potential of bioactive natural products with confidence, precision, and scientific rigor.
Online Inquiry
Frequently Asked Questions (FAQ)
Q1: Which method should I choose?
Q2: Can these methods be applied to complex natural product extracts?
A: Yes. Several of our biophysical approaches are compatible with complex mixtures, provided appropriate experimental design and controls are used.