Thermostability
Assess protein thermal stability and melting temperatures for developability analysis
Thermostability
Thermostability assays measure how proteins respond to increasing temperatures, providing critical developability data for protein therapeutics and industrial applications. These measurements help identify variants with improved stability profiles and predict behavior under storage and processing conditions.
nanoDSF Technology
Our thermostability measurements use nanoDifferential Scanning Fluorimetry (nanoDSF), a highly sensitive technique that monitors protein unfolding through changes in intrinsic tryptophan and tyrosine fluorescence.
Sample Preparation
Purified proteins are prepared in standard buffer conditions or customer-specified formulations.
Temperature Ramping
Samples are subjected to controlled temperature increases while monitoring fluorescence changes.
Melting Point Determination
The midpoint of the unfolding transition (Tm) is determined from the fluorescence data.
Key Measurements
Primary Stability Metrics
Units: °C Meaning: Temperature at which 50% of proteins are unfolded
The most commonly reported stability metric, representing the temperature at the midpoint of the unfolding transition.
Typical ranges:
- High stability: >70°C
- Good stability: 50-70°C
- Moderate stability: 40-50°C
- Poor stability: <40°C
Units: °C Meaning: Temperature at which 50% of proteins are unfolded
The most commonly reported stability metric, representing the temperature at the midpoint of the unfolding transition.
Typical ranges:
- High stability: >70°C
- Good stability: 50-70°C
- Moderate stability: 40-50°C
- Poor stability: <40°C
Units: °C Meaning: Temperature where unfolding begins
The temperature at which proteins first start to unfold, providing insight into the lower limit of thermal stability.
Applications:
- Storage condition optimization
- Process temperature limits
- Formulation stability assessment
Units: °C Meaning: Temperature range of unfolding
Indicates how cooperative the unfolding process is - narrow transitions suggest cooperative unfolding, while broad transitions may indicate multiple domains or intermediates.
Advanced Analysis
Unfolding Kinetics
- Rate of unfolding at different temperatures
- Activation energy calculations
- Predictive modeling of stability
Multi-Domain Analysis
- Multiple transition identification
- Domain-specific stability assessment
- Cooperative vs. independent unfolding
Applications
Developability Assessment
Evaluate protein stability for pharmaceutical development, identifying variants suitable for therapeutic applications.
Formulation Optimization
Test protein stability under different buffer conditions, pH levels, and additive concentrations.
Stability Engineering
Guide protein engineering efforts to improve thermal stability through rational or evolutionary approaches.
Quality Control
Monitor protein stability during development, manufacturing, and storage processes.
Experimental Conditions
Standard Conditions
Buffer Systems
- PBS (pH 7.4) - Standard physiological conditions
- Histidine buffer (pH 6.0) - Common formulation buffer
- Custom buffers - Customer-specified conditions
Temperature Range
- Typical: 20°C to 95°C
- Customizable based on protein characteristics
- High-resolution scanning (0.3°C/min)
Custom Formulations
pH Studies
- Multiple pH conditions (4.0-8.0)
- Buffer system optimization
- pH stability profiling
Additive Screening
- Stabilizing agents (trehalose, sucrose, arginine)
- Destabilizing conditions (urea, guanidine)
- Excipient compatibility testing
Data Analysis
Curve Fitting
Transition Modeling
- Two-state unfolding models
- Multi-transition analysis for complex proteins
- Thermodynamic parameter extraction
Statistical Analysis
- Replicate analysis and error estimation
- Comparative statistics between variants
- Quality control metrics
Comparative Analysis
Ranking and Selection
- Tm-based variant ranking
- Multi-parameter optimization
- Structure-stability relationships
Predictive Modeling
- Stability under storage conditions
- Accelerated stability predictions
- Formulation stability forecasting
Integration with Other Assays
Thermostability measurements complement other characterization assays:
Binding-Stability Relationships
- Correlation between binding affinity and thermal stability
- Trade-off analysis for optimization decisions
- Stability-function balance assessment
Developability Profiles
- Combined analysis with aggregation propensity
- Viscosity and stability correlations
- Comprehensive developability scoring
When to Use Thermostability Assays
Therapeutic Development
Therapeutic Development
Essential for: Protein therapeutics requiring long-term storage stability
Provides critical data for regulatory submissions and formulation development. Helps identify variants that can withstand manufacturing processes and maintain stability during storage.
Protein Engineering
Protein Engineering
Valuable for: Optimization campaigns focused on improving protein stability
Guides engineering efforts by providing quantitative feedback on stabilizing mutations. Essential for understanding structure-stability relationships.
Formulation Development
Formulation Development
Important for: Optimizing storage and delivery conditions
Tests protein behavior under different formulation conditions to identify optimal storage and delivery formats.
Industrial Applications
Industrial Applications
Critical for: Proteins used in industrial processes
Ensures proteins can withstand process conditions and maintain activity under operational temperatures.
Quality Control
Measurement Validation
Technical Replicates
- Multiple measurements per sample
- Statistical analysis of reproducibility
- Quality metrics for each measurement
Reference Standards
- Known stability controls
- Instrument performance verification
- Batch-to-batch consistency monitoring
Data Quality Assessment
Signal Quality
- Fluorescence signal intensity validation
- Baseline stability assessment
- Transition clarity evaluation
Fit Quality
- Goodness of fit statistics
- Residual analysis
- Parameter confidence intervals
Results Package
Your thermostability results include:
Quantitative Data
- Melting temperatures (Tm) with confidence intervals
- Onset temperatures and transition widths
- Comparative ranking of all variants tested
Visual Analysis
- Melting curves showing fluorescence vs. temperature
- Derivative plots highlighting transition points
- Comparative overlays for easy variant comparison
Analysis Reports
- Stability assessment with recommendations
- Structure-stability correlations when structural data available
- Formulation recommendations based on stability profiles
Getting Started
Ready to assess protein thermostability?
- Select your protein variants for stability assessment
- Choose experimental conditions (standard or custom buffers)
- Configure your experiment through the Foundry Portal
- Review conditions with our technical team
Thermostability assays are particularly valuable when combined with binding measurements. Consider running both assays to understand binding-stability trade-offs in your optimization campaigns.