Construction of a versatile expression library for all human single-pass transmembrane proteins for receptor pairings by high throughput screening.

•Interactions between protein ligands and receptors play crucial roles in cell-cell signaling.•Most of the human cell surface receptors have been identified but many of their corresponding ligands remain unknown.•We described a unique design of Fab fusion for the construction an expression library with versatile applications in HTP screening paradigms.•We constructed the expression library which has superior success rate of protein production.•We screened the library against VISTA, an important immune-checkpoint regulator, and identified IgSF11 as a novel binding partner of VISTA.