Increasing the Sensitivity of Enzyme-Linked Immunosorbent Assay Using Multiplexed Electrokinetic Concentrator
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Han_Increasing the sensitivity.pdf
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Author(s)
Cheow, Lih Feng
Ko, Sung Hee
Kim, Sung Jae
Kang, Kwan Hyoung
Han, Jongyoon
Alternative Title
Increasing the Sensitivity of ELISA using Multiplexed
Electrokinetic Concentrator
Electrokinetic Concentrator
Date Issued
March 2010
Journal
Analytical Chemistry
Publisher
American Chemical Society (ACS)
Citation
Cheow, Lih Feng et al. “Increasing the Sensitivity of Enzyme-Linked Immunosorbent Assay Using Multiplexed Electrokinetic Concentrator.” Analytical Chemistry 82.8 (2010): 3383–3388.
Version
Author's final manuscript
Abstract
We developed a novel method to increase the sensitivity of standard enzyme-linked immunosorbent assay (ELISA) using a multiplexed electrokinetic concentration chip. The poly(dimethylsiloxane) (PDMS) molecular concentrator(1) was used to trap and collect charged fluorescent product of target-bound enzyme turnover reaction of ELISA that occurred in a standard 96 well plate. Detection sensitivities of both prostate specific antigen (PSA) and CA 19-9 (a human pancreatic and gastrointestinal cancer marker) ELISAs in serum are enhanced ~100 fold with a low CV of <17%. We also integrated this method with an on-chip bead-based ELISA that lends itself toward a fully automated on-chip diagnostic device. Detection sensitivity of microfluidic bead-based CA 19-9 ELISA in serum is enhanced ~65 fold compared to the results without the electrokinetic accumulation step. This chip can be directly applied to enhance the readout sensitivity of a wide range of existing ELISA kits at concentrations below the current detection limit.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
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Creative Commons Attribution-Noncommercial-Share Alike 3.0
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DOI of Published Version
https://doi.org/10.1021/ac9024335
