Aptamer-linked assay for thrombin using gold nanoparticle amplification and inductively coupled plasma-mass spectrometry detection.
We describe a sensitive and specific sandwich assay for human alpha-thrombin. The assay takes advantage of sandwich binding of two affinity aptamers for increased specificity, gold nanoparticles for signal amplification, magnetic beads for fast magnetic separation, and inductively coupled plasma mass spectrometry for ultrasensitive detection. Other proteins, such as immunoglobulin G, serum albumin, transferrin, fibrinogen, and lysozyme did not show interference with the assay for human alpha-thrombin. The detection limit of human alpha-thrombin was as low as 0.5 fmol, corresponding to 10 pM thrombin in 50 microL, and the dynamic range covered approximately 3 orders of magnitude.
Zhao
Qiang
Q
Division of Analytical and Environmental Toxicology, Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB, Canada T6G 2G3.
Lu
Xiufen
X
Yuan
Chun-Gang
CG
Li
Xing-Fang
XF
Le
X Chris
XC
D052157:Aptamers, Nucleotide / Q000378:metabolism*; D015374:Biosensing Techniques / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D008280:Magnetics; D013058:Mass Spectrometry / Q000379:methods; D053768:Metal Nanoparticles / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013917:Thrombin / Q000032:analysis* / Q000378:metabolism
D052157:Aptamers, Nucleotide; D006046:Gold; D013917:Thrombin