NanoDSF is an advanced differential scanning fluorimetry method to measure the thermal stability of protein under a range of conditions, including through the binding of small-molecule ligands. Creative Biolabs has established excellent platforms for drug development. We provide s a variety of biophysical characterization services to meet the diverse needs of our customers.
Description of the NanoDSF Technology
NanoDSF is usually performed by using a real-time PCR instrument. In NanoDSF, a protein in solution is exposed to a temperature gradient that will lead to the unfolding of the protein. The intrinsic fluorescence of the protein, mainly originating from the aromatic side-chains of tyrosine and tryptophan residues, is examined. Upon unfolding, the environment of these residues will alter because they become exposed to the solvent and thus, their fluorescence intensity will change. The relation between fluorescence intensity changes and temperature gradient can be used to obtain a so-called apparent melting temperature (Tm). Similarly, information can be gained indicating cooperative (two-state) or complex unfolding transitions in multi-domain proteins.
Fig.1 Typical recording of fluorescence intensity versus temperature for the unfolding of protein.
Services
NanoDSF technologies have been optimized by Creative Biolabs for high-throughput screening applications, including chemical profiling of different protein families, identifying novel ligands, and investigating the stability of large numbers of proteins in buffers with different chemical compositions.
NanoDSF technology is specifically tailored for thermal and chemical unfolding studies. By monitoring fluorescence shifts of the amino acid tryptophan or tyrosine, unfolding profiles of multidomain proteins, transmembrane proteins or low-abundant drug targets can be obtained in an entirely label-free approach. For example, NanoDSF has been applied to the screening of monoclonal antibody formulations. Antibodies were screened in many different buffers, and the elevated aggregation levels induced by salt, pH, and high protein concentrations were successfully assessed.
The NanoDSF technology is a general method for the identification of ligands of target proteins from compound libraries. Using intrinsic fluorescence to monitor protein thermal unfolding, the ligand-binding affinity can be assessed from the shift of the unfolding temperature obtained in the presence of ligands relative to that obtained in the absence of ligands.
Advantages of NanoDSF
Creative Biolabs is one of the world’s leading service providers for biophysics-based assays. Our state-of-the-art portfolio of biophysical technologies enables us to provide you with outstanding support. If you are interested in our services, please contact us for more details.
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