Conditional protein-protein interactions tuned toward selective enzyme inhibition for use in loop-mediated isothermal amplification (LAMP) reactions
Isothermal amplification is a key technology for point-of-care diagnostics, but LAMP polymerases often exhibit significant off-target amplification. While hot-start inhibitors are widely used for thermostable enzymes, heat-labile polymerases like Bst require more selective control strategies. Here, we introduce a novel approach using camelid single-domain antibodies (VHH) as conditional inhibitors to improve LAMP assay specificity, with promising implications for next-generation diagnostics.
Using antibody discovery methods—including immunization, phage display library construction, affinity panning, and ELISA screening—we isolated VHH domains that partially or fully inhibit Bst polymerase catalysis. Screening workflows enabled selection of a lead VHH candidate that reduces off-target activity with minimal impact on processivity.
