Abstract
White spot disease (WSD) caused by white spot syndrome virus (WSSV) is one of the most dangerous diseases in the shrimp production industry, but there is no effective drug to control WSSV infection to date. The main challenges are the existence of biological barriers and untargeted drug delivery. In this study, a single-chain fragment variable (scFv) antibody-mediated WSSV-targeting drug delivery system was constructed for WSD targeting therapy. Natural compound geniposidic acid (GPA) could directly inhibit the expression of WSSV structural proteins VP24, VP26, and VP28 in Drosophila S2 cells. Yeast two-hybrid and intracellular co-localization showed that scFv antibody P75E8 could recognize and bind to VP28. Indirect ELISA showed that P75E8 had a good affinity to WSSV. Moreover, single-walled carbon nanotubes, GPA, and P75E8 were respectively selected as the nanocarrier, anti-WSSV drug, and targeting ligand to construct targeted drug delivery system SWCNTs-GPA-P75E8. Compared with GPA, SWCNTs-GPA-P75E8 showed a stronger anti-WSSV activity by inhibiting WSSV replication in crayfish and reducing the mortality of WSSV-infected crayfish. The mortality of WSSV-infected crayfish decreased by 23.33% and 46.67% after treatment with GPA and SWCNTs-GPA-P75E8 for 240 h, respectively. The results of GPA metabolism showed that SWCNTs-GPA-P75E8 could transport more GPA to target tissues of WSSV. Taken together, SWCNTs-GPA-P75E8 showed stronger WSSV-targeting ability and anti-WSSV activity. WSSV-specific antibody-mediated targeted drug delivery system is an effective strategy for WSD targeting therapy.
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