ArticleFrontiers in microbiology2016
Development of Monoclonal Antibody and Diagnostic Test for Middle East Respiratory Syndrome Coronavirus Using Cell-Free Synthesized Nucleocapsid Antigen.
Article in Frontiers in microbiology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed, 38 citations in OpenAlex.
- Eukaryotic cell-free protein synthesis: Chassis diversification, system engineering, and emerging applications.Synthetic and systems biotechnology · 2026Review
- On-site detection of MERS-CoV infections in a camel slaughterhouse in Kenya using a commercial rapid antigen test.Frontiers in veterinary science · 2025Article
- Applications of the wheat germ cell-free protein synthesis system in plant biochemical studies.Plant biotechnology (Tokyo, Japan) · 2024Article
- Review
- Generation and Utilization of a Monoclonal Antibody against Hepatitis B Virus Core Protein for a Comprehensive Interactome Analysis.Microorganisms · 2022Article
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- Recent advances in immunoassay technologies for the detection of human coronavirus infections.Frontiers in cellular and infection microbiology · 2022Review
- Review detection of Newcastle disease virus.Frontiers in veterinary science · 2022Review
- All-Trans Retinoic Acid Exhibits Antiviral Effect against SARS-CoV-2 by Inhibiting 3CLpro Activity.Viruses · 2021Article
- Highly specific monoclonal antibodies and epitope identification against SARS-CoV-2 nucleocapsid protein for antigen detection tests.Cell reports. Medicine · 2021Article
- Article
- Easy Synthesis of Complex Biomolecular Assemblies: Wheat Germ Cell-Free Protein Expression in Structural Biology.Frontiers in molecular biosciences · 2021Review
- Development of Monoclonal Antibodies and Antigen-Capture ELISA for Human Parechovirus Type 3.Microorganisms · 2020Article
- Development of SARS-CoV-2 Nucleocapsid Specific Monoclonal Antibodies.bioRxiv : the preprint server for biology · 2020Article
- An update on antiviral antibody-based biopharmaceuticals.International immunopharmacology · 2020Review
- Built-in RNA-mediated chaperone (chaperna) for antigen folding tailored to immunized hosts.Biotechnology and bioengineering · 2020Article
- A novel luciferase immunosorbent assay performs better than a commercial enzyme-linked immunosorbent assay to detect MERS-CoV specific IgG in humans and animals.Biosafety and health · 2019Article
- Rapid multiplex microfiber-based immunoassay for anti-MERS-CoV antibody detection.Sensing and Bio-Sensing Research · 2019Article
- Molecular Characteristics, Functions, and Related Pathogenicity of MERS-CoV Proteins.Engineering (Beijing, China) · 2019Review
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Authors and funding
10 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Protein nativity is one of the most critical factors for the quality of antigens used as immunogens and the reactivities of the resultant antibodies. The preparation and purification of native viral antigens in conventional cell-based protein expression systems are often accompanied by technical hardships. These challenges are attributable mainly to protein aggregation and insolubility during expression and purification, as well as to very low expression levels associated with the toxicity of some viral proteins. Here, we describe a novel approach for the production of monoclonal antibodies (mAbs) against nucleocapsid protein (NP) of the Middle East respiratory syndrome coronavirus (MERS-CoV). Using a wheat germ cell-free protein synthesis system, we successfully prepared large amounts of MERS-CoV NP antigen in a state that was highly soluble and intact for immunization. Following mouse immunization and hybridoma generation, we selected seven hybridoma clones that produced mAbs with exclusive reactivity against MERS-CoV NP. Epitope mapping and subsequent bioinformatic analysis revealed that these mAbs recognized epitopes located within relatively highly conserved regions of the MERS-CoV amino-acid sequence. Consistently, the mAbs exhibited no obvious cross-reactivity with NPs derived from other related viruses, including SARS coronavirus. After determining the optimal combinations of these mAbs, we developed an enzyme-linked immunosorbent assay and a rapid immunochromatographic antigen detection test that can be reliably used for laboratory diagnosis of MERS-CoV. Thus, this study provides strong evidence that the wheat germ cell-free system is useful for the production of diagnostic mAbs against emerging pathogens.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.