Complex integral membrane proteins with the property of multiple transmembrane-spanning regions are large protein families, including G-protein-coupled receptors (GPCR), ion channels, and transporters. Due to their essential roles in sensing and processing signals, they are the primary drug targets of more than half of the approved drugs—majorly small molecules. Although antibodies succeeded in the pharmaceutical markets, they are rare in modulating complex integral membrane proteins with favorable properties. Two significant limitations in such antibody discovery are the preparation of correctly folded antigens and the generation of antibodies against the natural conformation. Here, we developed an amphipol-trapped antigen as an immunogen and induced efficient mouse memory B cell responses. We generated antibodies unbiasedly by culturing single memory B cells and characterized their specificities. We implemented our strategy to generate high-affinity antibodies against the native conformation of vesicular monoamine transporter 2 (VMAT2), demonstrating the potential use in discovering antibodies against membrane proteins, including multi-pass transmembrane protein, for therapeutics.

 

Eighty-eight antibody sequences (heavy chain and light chains) discovered in this study using Sanger sequencing are included in this dataset. They were isolated from mouse memory B cells.