ArticleBiosensors & bioelectronics2018
Paper-based diagnostics in the antigen-depletion regime: High-density immobilization of rcSso7d-cellulose-binding domain fusion proteins for efficient target capture.
Article in Biosensors & bioelectronics, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- A colorimetric lateral flow immunoassay based on oriented antibody immobilization for sensitive detection of SARS-CoV-2.Sensors and actuators. B, Chemical · 2023Article
- Engineered Glycan-Binding Proteins for Recognition of the Thomsen-Friedenreich Antigen and Structurally Related Disaccharides.ACS chemical biology · 2023Article
- Nanobody-Functionalized Cellulose for Capturing SARS-CoV-2.Applied and environmental microbiology · 2022Article
- Generation of Thermally Stable Affinity Pairs for Sensitive, Specific Immunoassays.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Developing a SARS-CoV-2 Antigen Test Using Engineered Affinity Proteins.ChemRxiv : the preprint server for chemistry · 2021Article
- A rapid simple point-of-care assay for the detection of SARS-CoV-2 neutralizing antibodies.Communications medicine · 2021Article
- Emulsion Agglutination Assay for the Detection of Protein-Protein Interactions: An Optical Sensor for Zika Virus.ACS sensors · 2019Article
- Antimicrobial Resistance in the Asia Pacific region: a meeting report.Antimicrobial resistance and infection control · 2019Article
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Authors and funding
5 authors.
Funding
Abstract
In this work, we report the development of a general strategy for enhancing the efficiency of target capture in immunoassays, using a bifunctional fusion protein construct which incorporates a substrate-anchoring moiety for the high-abundance immobilization of an antigen-binding domain. This approach was informed by the development of a pseudo first-order rate constant model, and tested in a paper-based assay format using a fusion construct consisting of an rcSso7d binding module and a cellulose-binding domain. These rcSso7d-CBD fusion proteins were solubly expressed and purified from bacteria in high molar yields, and enable oriented, high-density adsorption of the rcSso7d binding species to unmodified cellulose within a 30-second incubation period. These findings were validated using two distinct, antigen-specific rcSso7d variants, which were isolated from a yeast surface display library via flow cytometry. Up to 1.6 micromoles of rcSso7d-CBD was found to adsorb per gram of cellulose, yielding a volume-averaged binder concentration of up to 760μM within the resulting active material. At this molar abundance, the target antigen is captured from solution with nearly 100% efficiency, maximizing the attainable sensitivity for any given diagnostic system.
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