Koutsoumpeli, Eleni orcid.org/0000-0002-3070-3319, Tiede, Christian, Murray, James et al. (4 more authors) (2017) Antibody mimetics for the detection of small organic compounds using a quartz crystal microbalance. Analytical Chemistry. pp. 3051-3058. ISSN 0003-2700
Abstract
Conventional immunoassays rely on antibodies that provide high affinity, specificity and selectivity against a target analyte. However, the use of antibodies for the detection of small-sized, non-immunogenic targets, such as pharmaceuticals and environmental contaminants presents a number of challenges. Recent advances in protein engineering have led to the emergence of antibody mimetics that offer the high affinity and specificity associated with antibodies but with reduced batch-to- batch variability, high stability and in vitro selection to ensure rapid discovery of binders against a wide range of targets. In this work we explore the potential of Affimers, a recent example of antibody mimetics, as suitable bio-receptors for the detection of small organic target compounds, here methylene blue. Target immobilisation for Affimer characterisation was achieved using long-chained alkanethiol linkers coupled with oligoethyleneglycol (LCAT-OEG). Using quartz crystal microbalance with dissipation monitoring (QCM-D), we determine the affinity constant, KD , of the methylene blue Affimer to be comparable to that of antibodies. Further, we demonstrate the high selectivity of Affimers for its target in complex matrices, here a limnetic sample. Finally, we demonstrate an Affimer-based competition assay, illustrating the potential of Affimers as bioreceptors in immunoassays for the detection of small-sized, non-immunogenic compounds.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2017, American Chemical Society. This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details. |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Electronic Engineering (York) |
Funding Information: | Funder Grant number EUROPEAN COMMISSION UNSPECIFIED |
Depositing User: | Pure (York) |
Date Deposited: | 08 Feb 2017 07:34 |
Last Modified: | 17 Dec 2024 00:07 |
Published Version: | https://doi.org/10.1021/acs.analchem.6b04790 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.1021/acs.analchem.6b04790 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:111982 |
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