Epitope mapping for monoclonal antibodies recognizing tuber necrotic isolates of potato virus y

Abstract

Potato virus Y (PVY) is an important viral pathogen of potato responsible for reducing tuber yield and quality across the globe. The PVYN and PVYNTN strains, the latter of which induces potato tuber necrotic ringspot disease (PTNRD), are regulated for international potato trade, and have been routinely detected using monoclonal antibodies (MAbs) that discriminate between PVYN and PVYO serotypes. Here, we identify the distinct binding sites in the capsid protein of PVY for three of the four main PVYN-specific MAbs, Bioreba-N, SASA-N, and Neogen-N, available commercially. These binding domains were mapped through a combination of TAS-ELISA testing of MAbs on multiple reference isolates of PVY, sequence analysis, heterologous expression of capsid protein fragments, and synthetic peptide binding experiments. All three MAbs were found to bind linear epitopes located within the first 31 N-terminal amino acids of the capsid protein. Bioreba-N MAb epitope spanned aa 1-17 and included three positions, aa 1, aa 11, and aa 17, which differ between PVYN and PVYO serotypes. Both SASA-N and Neogen-N epitopes spanned aa 22-30, and included two positions, aa 24 and aa 29, which differ between PVYN and PVYO serotypes. Epitopes for SASA-N and Neogen-N MAbs are likely to be identical or overlapping. Examination of available sequences for tuber necrotic isolates of PVY that do not react with PVYN-specific MAbs SASA-N and Neogen-N indicated possible selection for substitutions in corresponding epitopes leading to the loss of reactivity towards these antibodies. The data obtained suggested that testing with more than one PVYN serotype-specific MAb could assure a reliable serological identification of a PVYN or PVYNTN isolate.