title: Aptamer-based thrombin assay on microfluidic platform.
abstract: A facile and sensitive aptamer-based protocol has been developed for protein assay on microfluidic platform with fluorescence detection using an off-chip microarray scanner. Aptamer-functionalized magnetic beads were used to capture thrombin that binds to a second aptamer fluorescently labeled by Cy3. Experimental conditions, such as incubation time and temperature, washing time, interfering proteins, and aptamer, etc., were optimized for the microchip method. This work demonstrated there was a good relationship between fluorescence intensity and thrombin concentration in the range of 65-1000 ng/mL with the RSD less than 8%. Notably, an analysis only needs 1 μL volume of sample injection and this system can capture extremely tiny amount thrombin (0.4 fmol). This method has been successfully applied to assay of thrombin in human serum with the recovery of 79.74-95.94%.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering and Center of Materials Analysis, Nanjing University, Nanjing, P. R. China.
authors.forename: Fang-Yuan
authors.initials: FY
authors.lastname: Chen
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.forename: Hong-Zhen
authors.initials: HZ
authors.lastname: Lian
authors.forename: Hong-Yuan
authors.initials: HY
authors.lastname: Chen
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D016014:Linear Models; D059346:Magnets; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D008863:Microspheres; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013917:Thrombin / Q000032:analysis; D013997:Time Factors
chemical_list: D052157:Aptamers, Nucleotide; C081676:thrombin aptamer; D013917:Thrombin
keywords: Aptamer; Magnetic beads; Microarray scanner; Microfluidic chip; Thrombin