Field
Value
Language
dc.contributor.author
Shi, Lu
datacite.creator.affiliationIdentifier
https://ror.org/01aff2v68
en_US
datacite.creator.affiliation
University of Waterloo
en_US
datacite.creator.nameIdentifier
en_US
dc.contributor.author
Jin, Yan
datacite.creator.affiliationIdentifier
https://ror.org/0170z8493
en_US
datacite.creator.affiliation
Shaanxi Normal University
en_US
datacite.creator.nameIdentifier
en_US
dc.contributor.author
Liu, Juewen
datacite.creator.affiliationIdentifier
en_US
datacite.creator.affiliation
liujw@uwaterloo.ca
en_US
datacite.creator.nameIdentifier
https://orcid.org/0000-0001-5918-9336
en_US
dc.coverage.temporal
2023-06-17/2023-09-17
dc.date.accessioned
2023-10-04T16:04:31Z
dc.date.available
2023-10-04T16:04:31Z
dc.date.issued
2023-10-04
dc.identifier.uri
https://www.frdr-dfdr.ca/repo/dataset/b29adb15-b6f5-4785-a4fd-c02ed8430680
dc.identifier.uri
https://doi.org/10.20383/103.0817
dc.description
Lysozyme is a highly popular protein target for the development of aptamer-based biosensors. Because lysozyme is a polycation and DNA is a polyanion, it is essential to separate the contribution of nonspecific electrostatic interactions from specific aptamer binding. In this study, various factors affecting the binding of DNA and lysozyme, including DNA sequence, DNA length, pH, and salt concentration, were explored using fluorescence polarization. We concluded that direct fluorescence polarization and fluorescence intensity change are unlikely to be directly applicable for aptamer-based biosensors to detect lysozyme because all tested DNA sequences showed binding. These fundamental studies confirm the dominant role of electrostatic binding. We further evaluated three other methods including label-free fluorescent detection using a DNA staining dye, label-free colorimetric detection using gold nanoparticles, and a fluorescent sensor based on the strand displacement reaction. In each case, we focused on a random DNA sequence that is not expected to bind to lysozyme as an aptamer. Of all the methods, only the strand displacement strategy can be potentially used to evaluate aptamer binding, as the other methods all responded to non-aptamer sequences. This study provides valuable insights for assaying aptamer binding to cationic proteins that can exhibit nonspecific attraction to DNA.
en_US
dc.publisher
Federated Research Data Repository / dépôt fédéré de données de recherche
dc.rights
Creative Commons Public Domain Dedication (CC0 1.0)
en_US
dc.rights.uri
https://creativecommons.org/publicdomain/zero/1.0/
en_US
dc.subject
lysozyme
en_US
dc.subject
aptamer
en_US
dc.subject
Electrostatic-mediated Binding
en_US
dc.title
Electrostatic-mediated Binding of DNA to Lysozyme: Evaluation of Aptamer-based Assays for Highly Positively Charged Targets
en_US
globus.shared_endpoint.name
f163c1b3-9c88-42f6-a7bb-5839ed6c4063
globus.shared_endpoint.path
/2/published/publication_812/
datacite.publicationYear
2023
datacite.resourceType
Dataset
en_US
datacite.fundingReference.funderIdentifier
https://ror.org/01h531d29
en_US
datacite.fundingReference.funderName
Natural Sciences and Engineering Research Council of Canada
en_US
datacite.fundingReference.awardNumber
en_US
datacite.fundingReference.awardTitle
en_US
frdr.crdc.code
RDF1060101
en_US
frdr.crdc.group_en
Biological sciences
en_US
frdr.crdc.class_en
Biochemistry
en_US
frdr.crdc.field_en
Analytical biochemistry
en_US
frdr.crdc.group_fr
Sciences biologiques
fr_CA
frdr.crdc.class_fr
Biochimie
fr_CA
frdr.crdc.field_fr
Biochimie analytique
fr_CA
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