Next Article in Journal
Review of Transducer Principles for Label-Free Biomolecular Interaction Analysis
Previous Article in Journal
Nonlinear Impedance of Whole Cells Near an Electrode as a Probe of Mitochondrial Activity
Article Menu

Export Article

Open AccessArticle
Biosensors 2011, 1(2), 58-69; doi:10.3390/bios1020058

Kinetic ELISA in Microfluidic Channels

Department of Chemistry, University of Wyoming, Laramie, WY 82071, USA
*
Author to whom correspondence should be addressed.
Received: 26 April 2011 / Revised: 26 May 2011 / Accepted: 15 June 2011 / Published: 17 June 2011
View Full-Text   |   Download PDF [422 KB, uploaded 17 June 2011]   |  

Abstract

In this article, we describe the kinetic ELISA of Blue Tongue and Epizootic Hemorrhagic Disease viral antibodies in microfluidic channels by monitoring the rate of generation of the enzyme reaction product under static conditions. It has been shown that this format of the immunoassay allows very reliable quantitation of the target species using inexpensive glass microchips and a standard epifluorescence microscope system coupled to a CCD camera. For the viral antibodies assayed here, the limit of detection (LOD) for the analyte concentration in our microchips was established to be 3–5 times lower than that obtained on commercial microwell plates using a fiftieth of the sample volume and less than a third of the incubation time. Our analyses further show that when compared to the end-point ELISA format, the kinetic mode of this assay yields an improvement in the LOD by over an order of magnitude in microfluidic devices. This benefit is primarily realized as the observed variation in the background fluorescence (signal at the start of the enzyme reaction period) was significantly larger than that in the rate of signal generation upon repeating these assays in different microchannels/microchips. Because the kinetic ELISA results depend only on the latter quantity, the noise level in them was substantially lower compared to that in its end-point counterpart in which the absolute fluorescence measurements are of greater significance. While a similar benefit was also recorded through implementation of kinetic ELISAs on the microwell platform, the improvement in LOD registered in that system was not as significant as was observed in the case of microfluidic assays.
Keywords: microfluidic; ELISA; Blue Tongue Virus; Epizootic Hemorrhagic Disease Virus; viral antibodies; fluorescence microfluidic; ELISA; Blue Tongue Virus; Epizootic Hemorrhagic Disease Virus; viral antibodies; fluorescence
Figures

This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Yanagisawa, N.; Dutta, D. Kinetic ELISA in Microfluidic Channels. Biosensors 2011, 1, 58-69.

Show more citation formats Show less citations formats

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Biosensors EISSN 2079-6374 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top