ArticleBioconjugate chemistry2017
Bioorthogonal Protein Conjugation: Application to the Development of a Highly Sensitive Bioluminescent Immunoassay for the Detection of Interferon-γ.
Article in Bioconjugate chemistry, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 15 citations in OpenAlex.
- Novel Bioconjugate Materials: Synthesis, Characterization and Medical Applications.Advanced healthcare materials · 2025Review
- Characterization of microvessels in the human forehead dermis using intravascular dual perfusion and immunofluorescence staining.Scientific reports · 2025Article
- Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications.International journal of molecular sciences · 2020Review
- Bioluminescent detection of zearalenone using recombinant peptidomimetic Gaussia luciferase fusion protein.Mikrochimica acta · 2020Article
- The Smallest Isoform ofInternational journal of molecular sciences · 2020Article
- Bioluminescent Antibodies through Photoconjugation of Protein G-Luciferase Fusion Proteins.Bioconjugate chemistry · 2020Article
- Design of Gaussia luciferase-based bioluminescent stem-loop probe for sensitive detection of HIV-1 nucleic acids.The Analyst · 2018Article
Corrections and comments
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Authors and funding
9 authors at 2 institutions in 2 countries.
Funding
Abstract
Bioorthogonal conjugation eliminates the shortcomings of classical conjugation methods. The conjugation of antibodies to reporter proteins, such as bioluminescent protein, can be controlled with orthogonal conjugation methods. Here we report a bioluminescent immunoassay for the sensitive detection of interferon-γ (IFN-γ) that utilizes orthogonal conjugation of bioluminescent protein, Gaussia luciferase to anti-IFN-γ antibody. The IFN-γ is produced by the immune system and the detection of the IFN-γ is pivotal for the detection of persistent viral and bacterial infections. A bioorthogonal conjugation approach is used to conjugate an anti-IFN-γ antibody with a GLuc mutant containing the N-terminal tyrosine using formylbenzene diazonium hexafluorophosphate reagent (FBDP) in hydrophilic mild pH environment yielding high conjugation efficiency (60%). This reagent is shown to be specific for tyrosine (Tyr) residues. Therefore, conjugation through Tyr was orthogonal and not detrimental to the bioluminescence activity of GLuc. The immunoassay described in this paper is a sandwich type assay and involves a capture and a detection antibody. The assay was validated for its robustness, precision, accuracy, limit of detection, and recovery.
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