Flow Cytometry-Assisted Detection of Adenosine in Serum with an Immobilized Aptamer Sensor
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Date
2010-05-15
Authors
Huang, Po-Jung Jimmy
Liu, Juewen
Journal Title
Journal ISSN
Volume Title
Publisher
American Chemical Society
Abstract
Aptamers are single-stranded nucleic acids that can selectively bind to essentially any molecule of choice. Because of their high stability, low cost, ease of modification, and availability through selection, aptamers hold great promise in addressing key challenges in bioanalytical chemistry. In the past 15 years, many highly sensitive fluorescent aptamer sensors have been reported. However, few such sensors showed high performance in serum samples. Further challenges related to practical applications include detection in a very small sample volume and a low dependence of sensor performance on ionic strength. We report the immobilization of an aptamer sensor on a magnetic microparticle and the use of flow cytometry for detection. Flow cytometry allows the detection of individual particles in a capillary and can effectively reduce the light scattering effect of serum. Since DNA immobilization generated a highly negatively charged surface and caused an enrichment of counterions, the sensor performance showed a lower salt dependence. The detection limits for adenosine are determined to be 178 μM in buffer and 167 μM in 30% serum. Finally, we demonstrated that the detection can be carried out in 10 μL of 90% human blood serum.
Description
This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, copyright © American Chemical Society after peer review and technical editing by publisher. To access the final edited and published work see Huang, P.-J. J., & Liu, J. (2010). Flow Cytometry-Assisted Detection of Adenosine in Serum with an Immobilized Aptamer Sensor. Analytical Chemistry, 82(10), 4020–4026. https://doi.org/10.1021/ac9028505
Keywords
DNA, Adenosine, Aptamers