Differential Scanning Fluorescence (DSF) Technology

Differential scanning fluorescence (DSF) is a widely used technique to determine the apparent melting temperature (Tma) of purified proteins. CD Formulation applies cutting-edge DSF technology to our protein/peptide biopharmaceutical development and manufacturing services to help customers obtain more comprehensive and reliable information about the thermal stability of protein/peptide drugs.

What is Differential Scanning Fluorescence (DSF)?

DSF is a biophysical technique used to study the stability and thermodynamic properties of proteins in a specific environment, either under selected buffer conditions or in the presence of (partially) saturated target ligands. The technique uses molecular fluorescent dyes to monitor different conformations of proteins at different temperatures and assess their thermal stability.

Differential Scanning Fluorescence (DSF) Principle

DSF is a method for observing changes in protein fluorescence or absorbance when fluorescent or UV-absorbing dyes bind to proteins. In their natural state, proteins have hydrophobic groups hidden inside their structure. When the temperature rises, proteins transform into a denatured state, exposing hydrophobic groups. The exposed hydrophobic groups can bind to the fluorescent dye Sypro Orange and be excited to emit fluorescence. By measuring the changes in the fluorescence signal, the melting temperature and conformational change characteristics of the protein can be determined, thereby revealing the folding state and stability of the protein.

Fig. 1 Typical DSF profile of a protein sample. (Gao K, et al., 200)

Our Services Related to Differential Scanning Fluorescence (DSF)

With decades of experience in supporting protein/peptide biopharmaceutical development and manufacturing with DSF technology, our team of highly qualified experts can provide a range of DSF-related services to accelerate the implementation and success of your project. Our DSF technology supports all stages of your protein/peptide drug development and manufacturing - from early research to late-release testing under GMP.

To study protein stability and thermal denaturation, our highly qualified scientists use DSF technology to reflect the thermal stability of proteins/peptides by measuring fluorescence intensity. At different temperatures, the fluorescence intensity varies due to the different amounts of hydrophobic groups exposed by the protein, and a fluorescence-temperature curve can be plotted. Our DSF service provides precise insights into protein stability, ligand binding, and structural changes.

In addition, we also use NanoDSF technology for protein thermal stability analysis. Compared with conventional DSF, it is applicable to a wider range of protein samples. However, it should be noted that NanoDSF requires that the target protein must have intrinsic fluorescence, usually the presence of aromatic side chains such as tryptophan and tyrosine.

We provide the following DSF-related services, including but not limited to:

Screening Buffers for Formulation Studies

During protein/peptide pre-formulation studies, specific additives or buffer composition changes are needed to increase protein/peptide solubility and improve the thermal stability of the target protein to prevent protein unfolding or aggregation - even at low temperatures. Our scientists use DSF technology to monitor the effects of different buffers on the thermal stability of the target protein/peptide to screen for the best formulation.

Protein/Peptide Thermal Stability Studies

Thermal stability is an important property of protein/peptide therapeutics because it affects their physicochemical processing, pharmacodynamics, and pharmacokinetics. We use DSF technology to determine the temperature at which a protein begins to denature (melting temperature or Tm), as well as the stability of the protein over a range of temperatures and conditions. This information is important for understanding protein stability and folding and for developing stable protein formulations for drug development.

Identification of Low Molecular Weight Ligands Binding to Proteins

DSF is a rapid and inexpensive screening method for identifying low molecular weight ligands binding to proteins. We determine the thermodynamics of protein therapeutics by monitoring the change in protein melting temperature (Tm) in the presence and absence of ligands. This process provides valuable information about protein-ligand interactions, which helps to understand the mechanism of action of drugs and guide their development and optimization.

Workflow of Our Differential Scanning Fluorescence (DSF)

Fig.2 Workflow of differential scanning fluorescence. (CD Formulation)

Advantages of Our Differential Scanning Fluorescence (DSF) Technology

• Accurate and comprehensive data analysis.
• Wide temperature range.
• Small sample volume.
• High throughput measurement.
• Low cost and simple operation.

Advantages of Our NanoDSF Technology

• Nano DSF can analyze multiple samples simultaneously in less time than DSF.
• Nano DSF tests require much less sample and are not buffer-restricted.
• Simple to operate, no external dyes or sample handling required, completely label-free.
• A wide range of protein samples can be assayed from concentrations as high as 250 mg/ml to as low as 5 ug/ml.
• High throughput, reproducibility, and accuracy for simultaneous protein unfolding and aggregation detection.

Custom Differential Scanning Fluorescence (DSF) Services

Why Choose Our Differential Scanning Fluorescence (DSF) Technology?

• We have a team of experts with rich experience in performing DSF analytical method development and validation.
• We have accumulated decades of expertise and successful project experience using DSF technology to support protein/peptide biopharmaceutical development.
• Our laboratories are equipped with state-of-the-art equipment and technologies to support any type of protein characterization.
• We provide flexible experimental design and customized solutions.
• Fast turnaround time: detailed technical reports are provided within 5-7 days.

Publication

CD Formulation aims to provide a powerful analytical tool for characterizing the thermal stability of protein/peptide biologics in development and manufacturing. Please feel free to contact us to speak with our team of scientists and learn how our DSF technology can support your product development.

How It Works

STEP 1

Submit a service request describing your specific research or development need.

STEP 2

We'll email you to provide your quote and confirm order details if applicable.

STEP 3

Execute the project with real-time communication, and deliver the final report promptly.