In drug discovery and development, such as formulation optimization, it is a pre-requisite to biophysically characterize the proteins and obtain their meaningful and empirical information. Due to the complexity of protein systems, no single technique or approach is robust enough to achieve conclusive elucidation. Creative Biostructure provides a variety of sensitive MagHelix™ techniques and the most appropriate strategies for the biophysical characterization of proteins or biomolecules.
Our MagHelix™ analytical methods for biophysical characterization of biomolecules include but are not limited to:
Figure. Small angle X-ray scattering analysis of the peptide treatment.
With the development of genomes and large-scale expression and purification of recombinant proteins, it is critical to know about these newly proteins characterization to further understand their biochemical roles and to enable structure-function relationship studies. Many efforts have made and lead to the creation of various methods that have been proved to be effective in the biophysical protein characterization.
Among which, isothermal titration calorimetry (ITC) and surface plasmon resonance (SPR) are both thermodenaturation-based techniques and widely used methods to detect protein-ligand interaction. ITC can detect the complete thermodynamic parameters including affinity, enthalpy and stoichiometry while SPR is suitable for measuring binding afﬁnity and only requires little protein. There have been shown that the thermodynamic data obtained by SPR and ITC have good correlations. Each method can be selected accordingly.
Protein thermal shift assay (PTSA) is also a thermodenaturation assay to measure protein thermal stability and various approaches can be used for PTSA including reporter dyes, fluorescence wavelengths, FastPP, etc.
STD NMR is widely used for ligand screening and characterization of protein binding, from which values such as KD, the association and dissociation kinetics can be obtained. It is a robust method and it only requires small quantities of nonlabeled macromolecules.
Differential Scanning Calorimetry (DSC), a thermoanalytical technique, monitors heat effect by measuring the amount of energy that a sample absorbs or releases when it is heated or cooled. Its applications include studying oxidative stability, safety screening, determining the melting behavior of complex organic materials, etc.
BLI measures biomolecular interactions based on the principle of optical interferometry, allowing real-time monitoring.
Based on the full understanding of various techniques and the requirements of every client, scientists from Creative Biostructure select appropriate strategies to assist your research for protein characterization.
Timothy M. Ryan, et al. Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterization. PeerJ. 2013 May 7; 1:e73.