Journal Article Epublish
PubMed: 41953754 DOI: 10.1002/jex2.70118
Generation of Nano-Antibodies Targeting the SNMP1 Membrane Receptor of Spodoptera littoralis Using Extracellular Vesicles.
J Extracell Biol 5(4):e70118
Authors: Alexis Dogliani, Laura Fradale, Céline Valeille, Christian Cambillau, Emmanuelle Jacquin-Joly, Alain Roussel, Josiane Kassis-Sahyoun
Affiliations
- CNRS UMR7255 Laboratoire d'Ingénierie des Systèmes Macromoléculaires LISM Marseille France.
- Institute of Ecology and Environmental Sciences of Paris iEES-Paris INRAE Sorbonne Université, CNRS IRD UPEC Université Paris Cité Versailles France.
- CNRS UMR7255 Laboratoire d'Ingénierie des Systèmes Macromoléculaires LISM Marseille France. Institute of Ecology and Environmental Sciences of Paris iEES-Paris INRAE Sorbonne Université, CNRS IRD UPEC Université Paris Cité Versailles France.
Abstract
Handling membrane proteins (MPs) in their native environment is crucial for ensuring proper folding and stability. Extracellular vesicles (EVs) offer a superior alternative to detergents and artificial lipid environments. Expressing recombinant MPs on the surface of EVs allows for their display in a native context, making them ideal for llama immunization to generate nano-antibodies (Nabs) targeting MPs. In this study, we focused on the sensory neuron membrane protein 1 (SNMP1) from Spodoptera littoralis, a key player in transferring hydrophobic pheromones from pheromone-binding proteins to olfactory receptors, thereby triggering behavioral responses in moths. Targeting this protein with Nabs could provide a potential biocontrol strategy by limiting moth reproduction. Llama immunization was carried out using EVs displaying SNMP1 on their surface. A multi-layer approach was then employed, incorporating an anchor protein co-displayed with the target protein to specifically expose the EVs of interest for biopanning through phage display and selection via ELISA. Nabs against SNMP1 were successfully isolated, and their interactions with the membrane protein were characterized using bio-layer interferometry (BLI) with the same multi-layer approach. Furthermore, epitope binning by BLI aligned with predictions made by AlphaFold3.
MeSH
Extracellular Vesicles, Immunization, Antibodies, Multiple Chronic Conditions, Membrane Proteins, Pheromones, Environment, Membranes