CTPF : A Thermal Shift Assay platform at ICSN
The CTPF platform “Protein Thermostability Screening by Fluorescence” or “Protein Thermal Shift Assay” is a platform open to academics and industrialists for service activities or collaborations on research projects.
The Thermal Shift Assay (TSA) technique makes it possible to measure the variation of the denaturation temperature of a protein sample, depending on the experimental conditions, during a rapid temperature gradient. The fluorescent dye, Sypro Orange, is used to measure the exposure of the hydrophobic regions of the protein during its unfolding.
TSA Applications
The quality of protein preparations and the efficiency of purifications being essential for the development of biochemical and structural studies of proteins, the CTPF platform can very quickly provide essential informations for their characterization, by studying their denaturation profile.
The main applications of this technique are to improve the purification of proteins, to compare the stability of mutant proteins, to measure the interaction with substrates, but also the search and screening of ligands of interest. More recent developments also now allow us to study more complex assemblies such as viral particles on the platform.
The screening experiments are carried out on QPCR devices which can screen 96 or 384 conditions in 20 to 60 min. Each denaturation curve is obtained in volumes of 5 to 20 µl using generally 1 to 3 µg of protein per reaction. The platform has different types of QPCR machines and a pipetting robot for the preparation of reaction mixtures.
Infrastructure and Networks
ICSN-CNRS:
The platform belongs to the Institute of Chemistry of Natural Substances on the CNRS site of Gif-sur-Yvette. The platform premises are located on the 1st floor of Building 21 on campus.
PARIS-SACLAY:
The platform is referenced on the Plug in Labs site of the Université Paris-Saclay Platforms.
IBiSA:
The platform obtained the IBiSA label in 2018 as part of the creation of the Criblage@ParisSaclay (C@PS) platform, by partnering with the CCCHD (CEA), CIBI (ICSN-CNRS) and CIBLOT (Paris-Saclay University) platforms.
