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Microarrays as Model Biosensor Platforms to Investigate the Structure and Affinity of Aptamers
Joint Authors
Martin, Jennifer A.
Chushak, Yaroslav
Hagen, Joshua A.
Kelley-Loughnane, Nancy
Chávez, Jorge L.
Source
Issue
Vol. 2016, Issue 2016 (31 Dec. 2016), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2016-03-03
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
Immobilization of nucleic acid aptamer recognition elements selected free in solution onto the surface of biosensor platforms has proven challenging.
This study investigated the binding of multiple aptamer/target pairs immobilized on a commercially available microarray as a model system mimicking biosensor applications.
The results indicate a minimum distance (linker length) from the surface and thymine nucleobase linker provides reproducible binding across varying conditions.
An indirect labeling method, where the target was labeled with a biotin followed by a brief Cy3-streptavidin incubation, provided a higher signal-to-noise ratio and over two orders of magnitude improvement in limit of detection, compared to direct Cy3-protein labeling.
We also showed that the affinities of the aptamer/target interaction can change between direct and indirect labeling and conditions to optimize for the highest fluorescence intensity will increase the sensitivity of the assay but will not change the overall affinity.
Additionally, some sequences which did not initially bind demonstrated binding when conditions were optimized.
These results, in combination with studies demonstrating enhanced binding in nonselection buffers, provided insights into the structure and affinity of aptamers critical for biosensor applications and allowed for generalizations in starting conditions for researchers wishing to investigate aptamers on a microarray surface.
American Psychological Association (APA)
Martin, Jennifer A.& Chushak, Yaroslav& Chávez, Jorge L.& Hagen, Joshua A.& Kelley-Loughnane, Nancy. 2016. Microarrays as Model Biosensor Platforms to Investigate the Structure and Affinity of Aptamers. Journal of Nucleic Acids،Vol. 2016, no. 2016, pp.1-11.
https://search.emarefa.net/detail/BIM-1108986
Modern Language Association (MLA)
Martin, Jennifer A.…[et al.]. Microarrays as Model Biosensor Platforms to Investigate the Structure and Affinity of Aptamers. Journal of Nucleic Acids No. 2016 (2016), pp.1-11.
https://search.emarefa.net/detail/BIM-1108986
American Medical Association (AMA)
Martin, Jennifer A.& Chushak, Yaroslav& Chávez, Jorge L.& Hagen, Joshua A.& Kelley-Loughnane, Nancy. Microarrays as Model Biosensor Platforms to Investigate the Structure and Affinity of Aptamers. Journal of Nucleic Acids. 2016. Vol. 2016, no. 2016, pp.1-11.
https://search.emarefa.net/detail/BIM-1108986
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1108986