An emerging paradigm to develop analytical methods based on immobilized transmembrane proteins and its applications in drug discovery

The way to immobilize a protein onto a solid surface is the cornerstone for developing many bioanalytical assays which are applied in protein functional assay, protein-ligand interaction analysis and drug screening. Most immobilization approaches are established for soluble or cytosolic proteins, however, when it comes to transmembrane proteins (TMPs) like G protein-coupled receptors (GPCRs), they are challenged, and innovation is needed. In the last decade, site-specific immobilization strategies taking advantages of click chemistry and bioorthogonal conjugation have been developed, which have improved the sensitivity, selectivity, and repeatability of the protein based analytical methods. In this review, we focus on the immobilization of TMPs, because they constitute the largest pharmacological targets in clinics and serve as a paradigm to develop new analytical methods for bioanalysis. In particular, we summarize the progresses of site-specific immobilization made for establishing biosensors, bioassays, and chromatographic methods to pursue drug-protein interaction analysis and library screening. •Up-to-date overview of bioinspired site-specific covalent immobilization for transmembrane proteins (TMPs).•Outline practical guidelines for TMP immobilizations.•Summary of current analytical techniques for drug-TMP interactions with prerequisite of protein immobilization.•Recent advances in TMP-affinity chromatography for protein-ligand interactions and high throughput drug discovery.•Future directions for cutting-edge immobilizations and next-generation analytical methods.