Continuous Signal Enhancement for Sensitive Aptamer Affinity Probe Electrophoresis Assay Using Electrokinetic Concentration

We describe an electrokinetic concentration-enhanced aptamer affinity probe electrophoresis assay to achieve highly sensitive and quantitative detection of protein targets in a microfluidic device. The key weaknesses of aptamer as a binding agent (weak binding strength/fast target dissociation) were...

Full description

Bibliographic Details
Main Authors: Cheow, Lih Feng (Contributor), Han, Jongyoon (Contributor)
Other Authors: Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science (Contributor)
Format: Article
Language:English
Published: American Chemical Society, 2014-05-21T20:39:43Z.
Subjects:
Online Access:Get fulltext
LEADER 01911 am a22002293u 4500
001 87076
042 |a dc 
100 1 0 |a Cheow, Lih Feng  |e author 
100 1 0 |a Massachusetts Institute of Technology. Department of Biological Engineering  |e contributor 
100 1 0 |a Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science  |e contributor 
100 1 0 |a Cheow, Lih Feng  |e contributor 
100 1 0 |a Han, Jongyoon  |e contributor 
700 1 0 |a Han, Jongyoon  |e author 
245 0 0 |a Continuous Signal Enhancement for Sensitive Aptamer Affinity Probe Electrophoresis Assay Using Electrokinetic Concentration 
260 |b American Chemical Society,   |c 2014-05-21T20:39:43Z. 
856 |z Get fulltext  |u http://hdl.handle.net/1721.1/87076 
520 |a We describe an electrokinetic concentration-enhanced aptamer affinity probe electrophoresis assay to achieve highly sensitive and quantitative detection of protein targets in a microfluidic device. The key weaknesses of aptamer as a binding agent (weak binding strength/fast target dissociation) were counteracted by continuous injection of fresh sample while band-broadening phenomena were minimized due to self-focusing effects. With 30 min of continuous signal enhancement, we can detect 4.4 pM human immunoglobulin E (IgE) and 9 pM human immunodeficiency virus 1 reverse transcriptase (HIV-1 RT), which are among the lowest limits of detection (LOD) reported. IgE was detected in serum sample with a LOD of 39 pM due to nonspecific interactions between aptamers and serum proteins. The method presented in this paper also has broad applicability to improve sensitivities of various other mobility shift assays. 
520 |a Singapore-MIT Alliance (Singapore MIT Alliance-II CE programme) 
520 |a Singapore. Agency for Science, Technology and Research (Scholarship) 
546 |a en_US 
655 7 |a Article 
773 |t Analytical Chemistry