Nanobody-Functionalized Magnetic Beads for Enrichment of Foodborne Botulinum Toxin Type A Light Chain
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In this study, for the first time, nanobody conjugated beads were developed to enrich botulinum serotype A. We evaluated two types of nanobody A8-functionalized magnetic beads, both of which effectively captured and enriched BoNT/A from dilute solutions and complex protein mixtures, achieving a concentration increase of approximately 12- to 30-fold. The A8@N-beads showed higher binding capacity than the A8@S-beads, suggesting that our self-synthesized NH2 beads work well in this application. Our data serve as evidence that using different bead types with nanobody conjugation is a promising enrichment approach for further detection of Lc/A, particularly in applications involving low-abundance targets or complex sample matrices, such as food safety screening or clinical diagnostics.
Abstract
In this study, for the first time, nanobody conjugated beads were developed to enrich botulinum serotype A. We evaluated two types of nanobody A8-functionalized magnetic beads, both of which effectively captured and enriched BoNT/A from dilute solutions and complex protein mixtures, achieving a concentration increase of approximately 12- to 30-fold. The A8@N-beads showed higher binding capacity than the A8@S-beads, suggesting that our self-synthesized NH2 beads work well in this application. Our data serve as evidence that using different bead types with nanobody conjugation is a promising enrichment approach for further detection of Lc/A, particularly in applications involving low-abundance targets or complex sample matrices, such as food safety screening or clinical diagnostics.