ArticleBioconjugate chemistry2020
Bioluminescent Antibodies through Photoconjugation of Protein G-Luciferase Fusion Proteins.
Article in Bioconjugate chemistry, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 21 citations in OpenAlex.
- Article
- Single-Particle Functionality Imaging of Antibody-Conjugated Nanoparticles in Complex Media.ACS applied bio materials · 2023Article
- Protein A-Nanoluciferase fusion protein for generalized, sensitive detection of immunoglobulin G.Analytical biochemistry · 2023Article
- Bioluminescent RAPPID Sensors for the Single-Step Detection of Soluble Axl and Multiplex Analysis of Cell Surface Cancer Biomarkers.Analytical chemistry · 2022Article
- A Bioluminescent Sensor for Rapid Detection of PPEP-1, aSensors (Basel, Switzerland) · 2021Article
- Affinity-Based Methods for Site-Specific Conjugation of Antibodies.Bioconjugate chemistry · 2021Review
- A plug-and-play platform of ratiometric bioluminescent sensors for homogeneous immunoassays.Nature communications · 2021Article
- Recombinant Peptide Mimetic NanoLuc Tracer for Sensitive Immunodetection of Mycophenolic Acid.Analytical chemistry · 2021Article
- Paper-Based Immunosensors with Bio-Chemiluminescence Detection.Sensors (Basel, Switzerland) · 2021Review
- Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications.International journal of molecular sciences · 2020Review
- Expanding the Scope of Antibody Rebridging with New Pyridazinedione-TCO Constructs.Bioconjugate chemistry · 2020Article
Corrections and comments
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Bioluminescent antibodies represent attractive detection agents in both bioanalytical assays and imaging. Currently, their preparation relies on genetic fusion of luciferases to antibodies or nonspecific chemical conjugation strategies. Here, we report a generic method to generate well-defined covalent antibody-luciferase conjugates starting from commercially available monoclonal antibodies. Our approach uses fusion proteins consisting of the bright blue light-emitting luciferase NanoLuc (NL) and an Fc-binding protein domain (Gx) that can be photo-cross-linked to the antibody using UV light illumination. Green and red color variants were constructed by tight fusion of the NanoLuc with a green fluorescent acceptor domain and introduction of Cy3, respectively. To increase the already bright NanoLuc emission, tandem fusions were successfully developed in which the Gx domain is fused to two or three copies of the NanoLuc domain. The Gx-NL fusion proteins can be efficiently photo-cross-linked to all human immunoglobulin G (IgG) isotypes and most mammalian IgG's using 365 nm light, yielding antibodies with either one or two luciferase domains. The bioluminescent antibodies were successfully used in cell immunostaining and bioanalytical assays such as enzyme-linked immunosorbent assay (ELISA) and Western blotting.
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