Enzyme Stability Profiling Service

Enzyme Stability Profiling is a specialized analytical service that quantifies the robustness and resistance of an enzyme to various environmental and physical stressors, including heat, pH extremes, organic solvents, and prolonged storage. The service measures the enzyme's half-life (t1/2) and melting temperature (Tm) or denaturation temperature (Td) under different conditions. This fundamental data is essential for determining optimal storage, handling, and reaction conditions for both industrial biocatalysis and pharmaceutical assays.

CD Biosynsis offers a dedicated CRO service for Enzyme Stability Profiling, leveraging advanced thermal and chemical denaturation techniques such as Differential Scanning Fluorimetry (DSF, or ThermoFluor), Circular Dichroism (CD), and activity-based half-life assays. We provide rigorous, high-resolution data that quantifies the enzyme's resilience. Our platform is crucial for rational enzyme engineering projects aimed at enhancing thermostability, for optimizing industrial bioprocesses, and for ensuring the integrity and shelf-life of therapeutic enzyme products.

Highlights

Our platform delivers comprehensive stability metrics essential for robust experimental design and industrial viability.

• Thermal Stability Quantification: Precisely determine the enzyme's melting temperature (Tm) or denaturation temperature (Td) under various buffer conditions.
• Chemical Stability Profiling: Assess enzyme activity and structure retention in the presence of chaotropes, detergents, or organic solvents.
• Long-Term Storage Assessment: Measure activity half-life (t1/2) under different storage temperatures and formulations (e.g., lyophilized vs. liquid).
• Ligand-Induced Stabilization: Evaluate how substrates, cofactors, or inhibitors affect the enzyme's stability, providing mechanistic insight.

Applications

Stability profiling is a foundational step for commercialization, engineering, and formulation development:

Enzyme Engineering

           

Guiding site-directed mutagenesis to select mutations that significantly increase thermal or chemical stability for biocatalysis.

Bioprocess Optimization

Defining the maximum operational temperature, pH range, and solvent tolerance for large-scale industrial enzymatic reactions.

Pharmaceutical Formulation

Determining optimal buffer components and excipients to maximize the shelf-life and activity of therapeutic enzymes.

Quality Control and Comparability

Comparing the stability profile of batch-to-batch enzyme preparations or different expression hosts to ensure consistent quality.

Platform

Our Stability Profiling platform utilizes established biophysical techniques for accurate, high-resolution measurements.

Differential Scanning Fluorimetry (DSF)

High-throughput method using a fluorescent dye to monitor protein unfolding as temperature is increased, yielding the Tm value.

Circular Dichroism (CD) Spectroscopy

Monitoring changes in the secondary structure (alpha-helix, beta-sheet) upon thermal or chemical stress, providing high-fidelity structural stability data.

Activity Half-Life Assays

Incubating the enzyme under stress conditions and periodically measuring residual catalytic activity over time to determine t1/2.

Denaturant Gradient Experiments

Using urea or guanidinium chloride gradients to measure free energy of unfolding (Delta G), providing a thermodynamic measure of stability.

High-Throughput Buffer Screening

Rapidly screening the stabilizing effects of hundreds of different buffers, salts, and additives in a single experiment (e.g., using DSF).

Workflow

Our Stability Profiling follows a structured approach to characterize both structural and functional stability under various conditions:

• Assay Setup and Baseline: Validate the activity assay (if required) and establish the native baseline stability of the enzyme in a reference buffer.
• Thermal and pH Stress Testing: Determine the enzyme's Tm across a range of pH values and screen the effect of different salt concentrations.
• Chemical and Solvent Profiling: Test the enzyme's structural integrity and activity in the presence of relevant concentrations of organic solvents or detergents.
• Kinetic Inactivation (Half-Life Determination): Monitor the decay of enzyme activity over time at relevant storage or reaction temperatures (e.g., 4 C, 25 C, 37 C).
• Data Analysis and Recommendation: Calculate all stability metrics (Tm, t1/2, Delta G), compare conditions, and recommend the optimal formulation for maximum stability.

CD Biosynsis provides quantitative stability data that guides critical decisions in enzyme handling and commercial application. Every project includes:

• Thermal Stability Metrics: Melting temperature (Tm) values for all tested conditions.
• Functional Half-Life Data: Calculated t1/2 values at various temperatures or storage formulations.
• Structural Integrity Curves: High-quality thermal denaturation curves (DSF or CD data).
• Optimal Formulation Report: Recommendations for the most stable buffer, pH, and storage conditions.