Differential Scanning Fluorimetry (DSF)

Differential scanning fluorimetry (DSF) is a widely utilized method for examining the thermal stability of antibody drugs. Creative Proteomics employs differential scanning fluorimetry technology in our characterization service, delivering detailed solutions and data analysis to assist customers in acquiring information on antibody drug thermal stability.

Principles of Differential Scanning Fluorimetry

Differential scanning fluorimetry determines protein thermal stability through a fluorescent dye as an indicator. SYPRO Orange is the most frequently employed dye, environmentally sensitive and hydrophobic. As temperature rises, the protein unfolds, its hydrophobic part exposed, and the dye binds to the protein's hydrophobic area, causing fluorescence to increase. When a particular compound binds to the target protein, it stabilizes it, leading to an increase in the melting temperature of the protein.

Fig. 1 Differential scanning fluorimetry-based analysis of the thermal stability of antibody drugs. (Qin, K. et al., 2020)

Our DSF-Based Antibody Drugs Characterization Services

Differential scanning fluorimetry is a potent technique for characterizing antibody drugs. At Creative Proteomics, we employ DSF technology to ascertain several crucial parameters in the development and quality control of these therapeutic molecules. This enables our clients to accelerate their antibody drug development process. Our capabilities encompass the following essential areas:

Thermal stability testing

Thermal stability is a vital property of biotherapeutic antibodies, as it impacts their physicochemical process, pharmacodynamics, and pharmacokinetics. We utilize DSF technology to determine heat stability parameters of antibody drugs in high throughput, as well as evaluate the impact of mutations, pH, ligand binding, ionic strength, and other factors on heat stability. This process helps us isolate factors that can enhance heat stability. This critical information assists in optimizing the development of antibody drugs and clarifying their efficacy and shelf life.

Antibody-receptor affinity analysis

At Creative Proteomics, we determine the binding affinity and thermodynamics of antibody drugs by monitoring changes in protein melting temperature (Tm) in the presence of ligands. This process provides valuable information on antibody-ligand interactions. Our comprehensive data analysis results aid in understanding the mechanism of action of antibody drugs and guide their development optimization.

Advantages of Our Differential Scanning Fluorimetry Technology

• Minimal sample amounts, making it suitable even for limited or precious antibody samples.
• High throughput measurement.
• Wide temperature range.
• Low cost and ease of operation.
• Accurate, comprehensive data analysis.

Creative Proteomics offers characterization services based on DSF technology to accelerate your antibody drug development. We use state-of-the-art equipment and follow strict quality control measures to ensure the highest standards of data integrity and reproducibility. Contact us to learn more about our service and we will be happy to serve you.

Reference

1. Qin, K.; et al. Thermostability detection and optimization of glycoengineered antibodies and antibody-drug conjugates based on differential scanning flouremitry analysis. Bioorganic Chemistry. 2020, 94: 103391.