title: Highly sensitive and quantitative human thrombospondin-1 detection by an M55 aptasensor and clinical validation in patients with atherosclerotic disease.
abstract: Aptamer-based biosensors (aptasensor) are powerful tools for rapid and sensitive biomarker detection. In this study, we report a DNA aptamer probe evolved from cell-SELEX that can recognize thrombospondin-1 protein in human plasma samples. The KD value of the aptamer M55 binding to thrombospondin-1 was determined as 0.5 ± 0.2 μM with an R(2) of 0.9144. A horseradish peroxidase-linked short oligo was complementarily bound onto the 3' end of the aptamer sequence to facilitate the 'smart' design of an M55-aptasensor for quantifying thrombospondin-1 protein in plasma samples. The limit of detection was 6.96 fM. Thrombospondin-1 is a glycoprotein with multiple biological functions, including inflammation, platelet aggregation and endothelial cell apoptosis, and is involved in the pathology of atherosclerosis. In total, 118 plasma subjects were analyzed by using the aptasensor measurement with 1 μL sample volume and 5 min incubation time. The thrombospondin-1 concentrations in ST-Elevation Myocardial Infarction patients with severe atherosclerotic plaque burden were statistically significantly higher than in the healthy volunteers without atherosclerosis conditions, suggesting that thromboposnidn-1 is a potential plasma biomarker for atherosclerosis progression.
authors.affiliation: NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore 119077, Singapore.
authors.forename: Kaili
authors.initials: K
authors.lastname: Ji
authors.affiliation: Yong Loo Lin School of Medicine, National University of Singapore, Singapore 119228, Singapore.
authors.forename: Leonardo Pinto
authors.initials: LP
authors.lastname: de Carvalho
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research, 20 Biopolis Way, #06-01, Singapore 138668, Singapore.
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Bi
authors.affiliation: Yong Loo Lin School of Medicine, National University of Singapore, Singapore 119228, Singapore.
authors.forename: Aruni
authors.initials: A
authors.lastname: Seneviratnankn
authors.affiliation: Translational Molecular Imaging Group, Singapore Bioimaging Consortium, Agency for Science, Technology and Research, Singapore 138667, Singapore.
authors.forename: Kishore
authors.initials: K
authors.lastname: Bhakoo
authors.affiliation: National University Heart Centre, Singapore 119074, Singapore.
authors.forename: Mark
authors.initials: M
authors.lastname: Chan
authors.affiliation: Department of Chemistry and NUS Environmental Research Institute, National University of Singapore, Singapore 117543, Singapore; Shenzhen Engineering Laboratory for Eco-efficient Polysilicate Materials, Peking University Shenzhen Graduate School, Shenzhen, P.R.C. 518055. Electronic address: chmlifys@nus.edu.sg.
authors.forename: Sam Fong
authors.initials: SF
authors.lastname: Yau Li
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics*; D015374:Biosensing Techniques / Q000295:instrumentation*; D003324:Coronary Artery Disease / Q000097:blood* / Q000175:diagnosis*; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019700:Thrombospondin 1 / Q000097:blood* / Q000235:genetics
chemical_list: D052157:Aptamers, Nucleotide; D015342:DNA Probes; D019700:Thrombospondin 1
keywords: Aptamer; Aptasensor; Atherosclerosis; Thrombospondin