University of Notre Dame Background
Circular dichroism (CD) spectroscopy is a common approach to measure thermal stability to assess the peptide binding affinity, quantify interactions between peptides and class I major histocompatibility complex (MHC) proteins, and study the physical properties of peptide/MHC complexes. Quantifying the strength of the interactions is crucial in the identification of T cell epitopes and evaluating the structure of the peptides. CD is most frequently utilized by the structural immunology community, but it requires a large sample size which reduces throughput and limits the broader applications of the thermal stability measurements. Fortunately, a new, multifunctional methodology that increases the throughput was introduced at Notre Dame.
Researchers at the University of Notre Dame have developed a more adaptable and scalable technique called differential scanning fluorimetry (DSF) to evaluate the binding affinity in a high throughput manner. DSF requires substantially less sample and measurements can be performed in RT-PCR instruments, allowing for simultaneous analysis of multiple samples. DSF has a wide range of applications and can be used in high throughput assays, complementing other peptide-MHC binding assays used in screening, epitope discovery, and vaccine design. DSF is the superior technique in measuring peptide/MHC thermal stability and could potentially provide new insight into peptide-MHC interactions.
Differential scanning fluorimetry based assessments of the thermal and kinetic stability of peptide-MHC complexes. doi: 10.1016/j.jim.2016.02.016.
DSF requires less sample and is done in a high throughput manner.
High throughput assays
Complementing other peptide-MHC binding assays used in screening