Development of Nanobodies against Mal de Río Cuarto virus major viroplasm protein P9-1 for diagnostic sandwich ELISA and immunodetection
- Llauger, Gabriela
- Monti, Demian Esteban
- Adúriz Guerrero, Matías
- Romão, Ema
- Dumón, Analía Delina
- Mattio, Maria Fernanda
- Wigdorovitz, Andrés
- Muyldermans, Serge
- Vincke, Cécile
- Parreño, Gladys Viviana
- del Vas, Mariana
DOIAbstract
Mal de Río Cuarto virus (MRCV) is a member of the genus Fijivirus of the family Reoviridae that causes a devastating disease in maize and is persistently and propagatively transmitted by planthopper vectors. Virus replication and assembly occur within viroplasms formed by viral and host proteins. This work describes the isolation and characterization of llama-derived Nanobodies (Nbs) recognizing the major viral viroplasm component, P9-1. Specific Nbs were selected against recombinant P9-1, with affinities in the nanomolar range as measured by surface plasmon resonance. Three selected Nbs were fused to alkaline phosphatase and eGFP to develop a sandwich ELISA test which showed a high diagnostic sensitivity (99.12%, 95% CI 95.21–99.98) and sp...