Label-Free Reading of Microarray-Based Immunoassays with Surface Plasmon Resonance Imaging

A simple method is presented for patterning of protein antigens at a gold surface for use in surface plasmon resonance (SPR) imaging experiments. Microfluidic devices fabricated from poly(dimethylsiloxane) were used to flow various fluids over a gold substrate in spatially defined channels. This tec...

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Veröffentlicht in:Analytical chemistry (Washington) 2004-12, Vol.76 (24), p.7257-7262
Hauptverfasser: Kanda, Vishal, Kariuki, James K, Harrison, D. Jed, McDermott, Mark T
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container_issue 24
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container_title Analytical chemistry (Washington)
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creator Kanda, Vishal
Kariuki, James K
Harrison, D. Jed
McDermott, Mark T
description A simple method is presented for patterning of protein antigens at a gold surface for use in surface plasmon resonance (SPR) imaging experiments. Microfluidic devices fabricated from poly(dimethylsiloxane) were used to flow various fluids over a gold substrate in spatially defined channels. This technique was used to pattern the surface chemistry of the gold as well as to adsorb antigens from solution to the modified substrates. The resulting antigen arrays were probed with complementary antibodies in order to demonstrate the effectiveness of the patterning for antibody capture experiments. SPR imaging was used to aid in the optimization of array fabrication and to observe the interactions of unlabeled antibodies with these microarrays. This work presents a means of fabricating microarrays with controlled surface density of antigens. SPR imaging provides both quantitative and qualitative evaluation of antibody binding in a label free format.
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subjects Analytical chemistry
Animals
Antibodies - analysis
Antigens - chemistry
Biochemistry
Cattle
Chemistry
Dimethylpolysiloxanes - chemistry
Exact sciences and technology
General, instrumentation
Immunoassay
Immunoassay - methods
Immunoglobulin G - immunology
Microarray Analysis - instrumentation
Microarray Analysis - methods
Microfluidics - methods
Miscellaneous
Sensitivity and Specificity
Surface Plasmon Resonance - methods
title Label-Free Reading of Microarray-Based Immunoassays with Surface Plasmon Resonance Imaging
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