ArticleScientific reports2021
Development of Nanobodies against Mal de Río Cuarto virus major viroplasm protein P9-1 for diagnostic sandwich ELISA and immunodetection.
Article in Scientific reports, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 11 citations in OpenAlex.
- Engineered Nanobodies for early and accurate diagnosis of dengue virus infection.PLoS neglected tropical diseases · 2025Article
- The Use of Heterologous Antigens for Biopanning Enables the Selection of Broadly Neutralizing Nanobodies Against SARS-CoV-2.Antibodies (Basel, Switzerland) · 2025Article
- A novel PolyAr87-based cell transfection protocol for nanobody expression optimized via a targeted design of transfection approach.Frontiers in bioengineering and biotechnology · 2025Article
- Development of nanobodies against the coat protein of maize chlorotic mottle virus.FEBS open bio · 2024Article
- Development of an immunodiagnostic assay for the detection ofTurkish journal of biology = Turk biyoloji dergisi · 2024Article
- New Virus Diagnostic Approaches to Ensuring the Ongoing Plant Biosecurity of Aotearoa New Zealand.Viruses · 2023Review
- 14th International dsRNA Virus Symposium, Banff, Alberta, Canada, 10-14 October 2022.Virus research · 2023Article
- Nanobodies: a promising approach to treatment of viral diseases.Frontiers in immunology · 2023Review
- Research progress and applications of nanobody in human infectious diseases.Frontiers in pharmacology · 2022Review
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Authors and funding
11 authors at 5 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
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.
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