Antibody Thermal Stability Determination

Antibody thermal stability determination is to ascertain the stability of an antibody during the heating process, identify its denaturation temperature (Tm), i.e., the temperature point at which the antibody begins to lose structural stability, and evaluate its long-term stability at a specific temperature. The thermal stability of an antibody directly affects its biological activity and functionality, which is crucial for drug development, diagnostic reagents, and research applications.

 

Common Methods

1. Differential Scanning Calorimetry (DSC)

(1) Principle: It measures the heat flow difference between the sample and the reference during the heating process to determine the denaturation temperature of the antibody protein.

(2) Application: It accurately assesses the thermal stability and denaturation temperature of the antibody, suitable for antibody engineering and formulation development.

 

2. Circular Dichroism (CD) Analysis

(1) Principle: It uses the difference in circularly polarized light absorption of the protein at specific wavelengths to monitor changes in the secondary structure of the protein.

(2) Application: It assesses the structural changes of the antibody at different temperatures and understands its stability characteristics.

 

3. Surface Plasmon Resonance (SPR)

(1) Principle: It monitors the change in affinity of the antibody binding to a specific antigen to evaluate the effect of temperature on its functionality.

(2) Application: It analyzes the functional stability of the antibody under different temperature conditions.

 

4. Fluorescence Spectroscopy

(1) Principle: It monitors the changes in the structure of the protein by measuring the fluorescence intensity changes of the internal tryptophan and tyrosine residues of the protein.

(2) Application: It quickly assesses the thermal stability of the antibody, suitable for preliminary screening and comparing the stability of different antibodies or formulations.

 

5. Dynamic Light Scattering (DLS)

(1) Principle: It measures the changes in particle size distribution in the antibody solution to assess the effect of temperature on its aggregation state.

(2) Application: It understands the aggregation trend of the antibody under different temperature conditions.

 

Considerations for Experimental Design

1. Sample Preparation

Ensure the purity and concentration of the antibody sample are suitable for the selected analysis method.

 

2. Temperature Range

Select a temperature range that covers the potentially stable and unstable areas of the antibody.

 

3. Data Analysis

Use appropriate software and statistical methods to accurately interpret the experimental data and draw meaningful conclusions.