Thrombin detection using a piezoelectric aptamer-linked immunosorbent assay

• Published in: Analytical biochemistry Vol. 443; no. 1; pp. 97 - 103
• Main Authors: Collins, Cheryl M., Yui, Samuel, Roberts, Charles E.S., Kojic, Igor
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.12.2013
• Subjects: Acoustic Membrane MicroParticle (AMMP); Acoustics; antibodies; Aptamer; Aptamers, Nucleotide - chemistry; Base Sequence; Biosensing Techniques; blood serum; chemiluminescence; Detection; Diagnostic; Humans; Immunosorbents - chemistry; Molecular Sequence Data; oligonucleotides; radioimmunoassays; rapid methods; Reproducibility of Results; SELEX; SELEX Aptamer Technique; Sensitivity and Specificity; Serum - chemistry; Thrombin; Thrombin - analysis; ViBE; Vibration; western blot analysis; Western blotting; Acoustic Membrane MicroParticle (AMMP); Diagnostic; Thrombin; Aptamer; Detection; ViBE; SELEX
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2013.08.019

The development of diagnostic assays using highly targeted specific aptamers with existing detection platforms has been an endeavor with few opportunities until now. Many current commercially available diagnostic platforms make use of detection systems employing capture agents composed of modified antigen-specific antibodies coupled with a variety of detection modalities, including radioimmunoassays, fluorescence-based detection assays, electro/chemiluminescence assays, and immunoradiometric assays. In the studies presented here, a novel frequency-modulating technology from BioScale called Acoustic Membrane MicroParticle (AMMP) detection was used to demonstrate a sensitive and reproducible method of incorporating aptamers as capture and detection agents. The method provides a robust and rapid detection of thrombin in human serum while also eliminating the labor-intensive efforts of Western blot analysis and is not affected by the interfering substances found in serum that often affect optical-based detection systems. In addition, we have demonstrated, for the first time, the adaptation of the AMMP platform to exploit aptamers against a clinically relevant target. The AMMP platform is an ideal medium for using aptamers in commercial assay development for application in a clinical setting.