924 Harnessing VLPs & banodiscs to unlock antibody discovery for challenging membrane proteins
Year: 2026
Type: conference-abstract
Abstract: Background Multi-pass transmembrane proteins, including G protein-coupled receptors, ion channels, transporters, and immune receptor complexes, represent a large class of validated therapeutic targets. However, their dependence on a native membrane environment creates significant challenges for recombinant expression, structural stability, and functional characterization, limiting their utility in antibody discovery. We developed virus-like particle (VLP) and styrene maleic acid (SMA)-based nanodisc platforms to display full-length membrane proteins in native-like lipid bilayers for antibody generation, screening, and bioanalytical applications.Methods Full-length membrane proteins were displayed on VLPs generated in mammalian cells or incorporated into SMA-based nanodiscs using detergent-free extraction methods. Biophysical characterization included transmission electron microscopy, dynamic light scattering, high-performance liquid chromatography, and purity analysis. Antigen functionality was evaluated using enzyme-linked immunosorbent assay, surface plasmon resonance, and biolayer interferometry. VLP-displayed CXCR4 was used for mouse immunization and monoclonal antibody discovery. Additional studies assessed biotinylated VLPs and nanodiscs for phage panning applications and nanodisc-displayed T cell receptor (TCR)-CD3 complexes for antibody screening.Results VLPs and nanodiscs successfully displayed multiple membrane protein classes, including STEAP1, CXCR4, adenosine A2A receptor, GPRC5D, and TCR-CD3 complexes, while maintaining native conformations and robust binding activity. STEAP1 VLPs demonstrated high purity and specific antibody binding by enzyme-linked immunosorbent assay. A2A receptor nanodiscs exhibited nanomolar affinity interactions by surface plasmon resonance and biolayer interferometry. Immunization with CXCR4 VLPs generated functional antibodies, including a lead clone that demonstrated ligand-blocking activity comparable to a commercial anti-CXCR4 antibody. Biotinylated VLPs and nanodiscs produced strong binding signals in phage-panning compatible assays. Nanodisc-displayed TCR-CD3 complexes enabled high-affinity binding and screening of anti-CD3 antibodies, supporting their use in biologically relevant antibody discovery workflows.Conclusions VLP and SMA-based nanodisc platforms provide robust, biologically active presentation of challenging membrane protein targets in native-like conformations. These technologies support antibody discovery, immune receptor characterization, and assay development against therapeutically relevant membrane proteins. By enabling functional interrogation of previously difficult-to-access targets, VLPs and nanodiscs offer versatile tools for advancing next-generation immunotherapies.
Author: Maggie Gao
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,490
OpenAlex ID: W7225148106