Area-scaling of interferometric and fluorescent detection of protein on antibody microarrays

Xuefeng Wang; Ming Zhao; David D. Nolte

doi:10.1016/j.bios.2008.07.078

Area-scaling of interferometric and fluorescent detection of protein on antibody microarrays

Xuefeng Wang, Ming Zhao, David D. Nolte

Optical Sciences, College of

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

The sensitivity limits for an in-line interferometric (IL)/fluorescent (FL) dual-channel BioCD are established as a function of spatial averaging. Forward-phase and sandwich assays at 10 ng/ml were performed on large-scale antibody microarrays (6800-spot) and detected using in-line interferometry and fluorescence channels. The interferometric channel has an extrapolated label-free limit-of-detection (LOD) of 250 pg/ml in a forward-phase assay for which the fluorescent channel is inapplicable. In the sandwich assay, the extrapolated limit-of-detection is 70 pg/ml for the interferometric channel, and for the fluorescent channel it is 30 pg/ml. Intrinsic scale-free sensitivities for the detection channels are defined assuming spatially uncorrelated fluctuations with sensitivities of S′ = 13 pg/mm for interferometric detection and 7 pg/mm for sandwich fluorescent detection. The intrinsic sensitivities become weakly scale dependent in the presence of fractal spatial correlations among the antibody spots.

Original language	English (US)
Pages (from-to)	981-987
Number of pages	7
Journal	Biosensors and Bioelectronics
Volume	24
Issue number	4
DOIs	https://doi.org/10.1016/j.bios.2008.07.078
State	Published - Dec 1 2008

Keywords

Antibody microarray
BioCD
Fluorescence
Immunoassay
Interferometry
Label-free
Scalability

ASJC Scopus subject areas

Biotechnology
Biophysics
Biomedical Engineering
Electrochemistry

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title = "Area-scaling of interferometric and fluorescent detection of protein on antibody microarrays",

abstract = "The sensitivity limits for an in-line interferometric (IL)/fluorescent (FL) dual-channel BioCD are established as a function of spatial averaging. Forward-phase and sandwich assays at 10 ng/ml were performed on large-scale antibody microarrays (6800-spot) and detected using in-line interferometry and fluorescence channels. The interferometric channel has an extrapolated label-free limit-of-detection (LOD) of 250 pg/ml in a forward-phase assay for which the fluorescent channel is inapplicable. In the sandwich assay, the extrapolated limit-of-detection is 70 pg/ml for the interferometric channel, and for the fluorescent channel it is 30 pg/ml. Intrinsic scale-free sensitivities for the detection channels are defined assuming spatially uncorrelated fluctuations with sensitivities of S′ = 13 pg/mm for interferometric detection and 7 pg/mm for sandwich fluorescent detection. The intrinsic sensitivities become weakly scale dependent in the presence of fractal spatial correlations among the antibody spots.",

keywords = "Antibody microarray, BioCD, Fluorescence, Immunoassay, Interferometry, Label-free, Scalability",

author = "Xuefeng Wang and Ming Zhao and Nolte, {David D.}",

note = "Funding Information: This work was sponsored under grants from Quadraspec, Inc. and from the Indiana Economic Development Corporation through the Purdue Research Foundation. Copyright: Copyright 2009 Elsevier B.V., All rights reserved.",

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AU - Zhao, Ming

AU - Nolte, David D.

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PY - 2008/12/1

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N2 - The sensitivity limits for an in-line interferometric (IL)/fluorescent (FL) dual-channel BioCD are established as a function of spatial averaging. Forward-phase and sandwich assays at 10 ng/ml were performed on large-scale antibody microarrays (6800-spot) and detected using in-line interferometry and fluorescence channels. The interferometric channel has an extrapolated label-free limit-of-detection (LOD) of 250 pg/ml in a forward-phase assay for which the fluorescent channel is inapplicable. In the sandwich assay, the extrapolated limit-of-detection is 70 pg/ml for the interferometric channel, and for the fluorescent channel it is 30 pg/ml. Intrinsic scale-free sensitivities for the detection channels are defined assuming spatially uncorrelated fluctuations with sensitivities of S′ = 13 pg/mm for interferometric detection and 7 pg/mm for sandwich fluorescent detection. The intrinsic sensitivities become weakly scale dependent in the presence of fractal spatial correlations among the antibody spots.

AB - The sensitivity limits for an in-line interferometric (IL)/fluorescent (FL) dual-channel BioCD are established as a function of spatial averaging. Forward-phase and sandwich assays at 10 ng/ml were performed on large-scale antibody microarrays (6800-spot) and detected using in-line interferometry and fluorescence channels. The interferometric channel has an extrapolated label-free limit-of-detection (LOD) of 250 pg/ml in a forward-phase assay for which the fluorescent channel is inapplicable. In the sandwich assay, the extrapolated limit-of-detection is 70 pg/ml for the interferometric channel, and for the fluorescent channel it is 30 pg/ml. Intrinsic scale-free sensitivities for the detection channels are defined assuming spatially uncorrelated fluctuations with sensitivities of S′ = 13 pg/mm for interferometric detection and 7 pg/mm for sandwich fluorescent detection. The intrinsic sensitivities become weakly scale dependent in the presence of fractal spatial correlations among the antibody spots.

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KW - Immunoassay

KW - Interferometry

KW - Label-free

KW - Scalability

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Area-scaling of interferometric and fluorescent detection of protein on antibody microarrays

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Keywords

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