Ver ítem
- xmlui.general.dspace_homeCentros e Institutos de InvestigaciónCICVyA. Centro de Investigación en Ciencias Veterinarias y AgronómicasInstituto de BiotecnologíaArtículos científicosxmlui.ArtifactBrowser.ItemViewer.trail
- Inicio
- Centros e Institutos de Investigación
- CICVyA. Centro de Investigación en Ciencias Veterinarias y Agronómicas
- Instituto de Biotecnología
- Artículos científicos
- Ver ítem
Development of nanobodies against Mal de Río Cuarto virus major viroplasm protein P9‑1 for diagnostic sandwich ELISA and immunodetection
Resumen
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
[ver mas...]
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 specificity (100%, 95% CI 96.31–100) and a detection limit of 0.236 ng/ml. Interestingly, these Nanobodies recognized different P9-1 conformations and were successfully employed to detect P9-1 in pull-down assays of infected maize extracts. Finally, we demonstrated that fusions of the Nbs to eGFP and RFP allowed the immunodetection of virus present in phloem cells of leaf thin sections. The Nbs developed in this work will aid the study of MRCV epidemiology, assist maize breeding programs, and be valuable tools to boost fundamental research on viroplasm structure and maturation.
[Cerrar]
Autor
Llauger, Gabriela;
Monti, Demian Esteban;
Aduriz Guerrero, Matías;
Romão, Ema;
Dumon, Analia Delina;
Mattio, Maria Fernanda;
Wigdorovitz, Andres;
Muyldermans, Serge;
Vincke, Cécile;
Parreño, Viviana;
Del Vas, Mariana;
Fuente
Scientific Reports 11 : 20013 (Octubre 2021)
Fecha
2021-10
Editorial
Springer Nature
ISSN
2045-2322
Documentos Relacionados
Formato
pdf
Tipo de documento
artículo
Proyectos
(ver más)
INTA/2019-PD-E4-I081-001/2019-PD-E4-I081-001/AR./Generación de reactivos, desarrollo de metodologías, validación y acreditación de ensayos para el diagnóstico de patógenos vegetales.
Palabras Claves
Derechos de acceso
Abierto
Excepto donde se diga explicitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)