PD-1/PD-L1 binding studies using microscale thermophoresis.

ABSTRACT: The characterization of protein interactions has become essential in many fields of life science, especially drug discovery. Microscale thermophoresis (MST) is a powerful new method for the quantitative analysis of protein-protein interactions (PPIs) with low sample consumption. In addition, one of the major advantages of this technique is that no tedious purification step is necessary to access the protein of interest. Here, we describe a protocol using MST to determine the binding affinity of the PD-1/PD-L1 couple, which is involved in tumour escape processes, without purification of the target protein from cell lysates. The method requires the overexpression of fluorescent proteins in CHO-K1 cells and describes the optimal conditions for determining the dissociation constant. The protoc

SUBMITTER: Magnez R 

PROVIDER: S-EPMC5732298 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature