Development of an Electrochemical Biosensor for Tetrodotoxin Using Specific Binding Peptide on Polypyrrole/Au Nanoparticle-Modified Electrodes

The concern regarding tetrodotoxin (TTX), a highly hazardous marine neurotoxin found in puffer fish, has expanded beyond Asia due to the migration of puffer fish caused by the rise in global temperatures. This highlights the urgent need to develop fast yet reliable methods for detecting TTX. In this study, we developed a peptide-based potentiometric TTX sensor based on a polypyrrole/Au nanoparticle-modified carbon screen-printed electrode (PPy/AuNP SPE). The bioreceptor responsible for recognizing TTX is a specific binding peptide that was discovered through phage display technique. The phage-displayed peptide candidates were sorted based on frequency and similarity, and their binding affinity was subsequently assessed via phage enzyme-linked immunosorbent assay. The C-terminal of the specific binding peptide was then modified with cysteamine to facilitate its immobilization through Au-S bonding on the PPy/AuNP SPE platform, thereby constructing the TTX sensor. The sensing platform was prepared by successive electrodeposition of polypyrrole and AuNP onto the surface of carbon SPE as a substrate. Both materials play significant roles to improve the poor conductivity of carbon SPE and provide sufficient immobilization sites for TTX receptors, respectively. Finally, the PPy/AuNP TTX sensor demonstrated a detection limit of around 2.80 ppb with a detection range from 2 to 1000 ppb, making it a promising platform for rapid and reliable marine toxin detection.