tANCHOR: A novel mammalian cell surface peptide display system

Abstract

A novel tool for the presentation of peptides and small proteins on the surface of human cells has been developed. Our tANCHOR system utilizes tetraspanin anchors containing heterologous amino acid sequences inserted instead of the large extracellular loop. This technology allows a highly effective extracellular display of epitopes for antibody binding studies and many other potential applications. METHOD SUMMARY We demonstrate that tetraspanins are able to present heterologous antigens instead of their large extracellular loop on the cell surface by employing confocal laser scanning microscopy. Cell-based ELISA techniques were used to identify the most efficient tetraspanin anchors for displaying of peptides on the surface of human cells. Extracellular accessibility of antigens by antibodies was confirmed by scanning electron microscopy. Our approach was further used to study tetraspanin-anchored 2F5 antigen binding of a broadly HIV-1 neutralizing antibody.