Array-Based ELISAs for High-Throughput Analysis of Human Cytokines

In this report, we describe the development of a mini-array system suitable for high-throughput quantification of proteins. This mini-array is a multiplexed, sandwich-type ELISA that measures the concentration of seven different human cytokines— TNF-α, IFNα, IFNγ, IL-1α, IL-1β, IL-6, and IL-10—from...

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Main Authors: M.D. Moody, S.W. Van Arsdell, K.P. Murphy, S.F. Orencole, C. Burns
Format: Article
Language:English
Published: Taylor & Francis Group 2001-07-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/01311dd03
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author M.D. Moody
S.W. Van Arsdell
K.P. Murphy
S.F. Orencole
C. Burns
author_facet M.D. Moody
S.W. Van Arsdell
K.P. Murphy
S.F. Orencole
C. Burns
author_sort M.D. Moody
collection DOAJ
description In this report, we describe the development of a mini-array system suitable for high-throughput quantification of proteins. This mini-array is a multiplexed, sandwich-type ELISA that measures the concentration of seven different human cytokines— TNF-α, IFNα, IFNγ, IL-1α, IL-1β, IL-6, and IL-10—from a single sample in each well of a 96-well plate. The mini-array is produced by spotting monoclonal antibodies (mAbs) in a 3 × 3 pattern in the bottom of the wells of 96-well polystyrene plates. Cytokines that are captured by the arrayed mAbs are detected by using biotinylated mAbs, followed by the addition of a streptavidin-horseradish peroxidase (HRP) conjugate and a chemiluminescent substrate. The light produced from the HRP-catalyzed oxidation of the substrate is measured at each spot in the array by imaging the entire plate with a commercially available CCD camera. Here, we demonstrate that these 96-well-plate format mini-arrays have performance characteristics that make them suitable for the high-throughput screening of anti-inflammatory compounds.
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spelling doaj-art-a5c39ea74ed74babab091657b3455d712025-08-20T02:25:51ZengTaylor & Francis GroupBioTechniques0736-62051940-98182001-07-0131118619410.2144/01311dd03Array-Based ELISAs for High-Throughput Analysis of Human CytokinesM.D. Moody0S.W. Van Arsdell1K.P. Murphy2S.F. Orencole3C. Burns41Pierce Endogen, Woburn, MA, USA1Pierce Endogen, Woburn, MA, USA1Pierce Endogen, Woburn, MA, USA1Pierce Endogen, Woburn, MA, USA1Pierce Endogen, Woburn, MA, USAIn this report, we describe the development of a mini-array system suitable for high-throughput quantification of proteins. This mini-array is a multiplexed, sandwich-type ELISA that measures the concentration of seven different human cytokines— TNF-α, IFNα, IFNγ, IL-1α, IL-1β, IL-6, and IL-10—from a single sample in each well of a 96-well plate. The mini-array is produced by spotting monoclonal antibodies (mAbs) in a 3 × 3 pattern in the bottom of the wells of 96-well polystyrene plates. Cytokines that are captured by the arrayed mAbs are detected by using biotinylated mAbs, followed by the addition of a streptavidin-horseradish peroxidase (HRP) conjugate and a chemiluminescent substrate. The light produced from the HRP-catalyzed oxidation of the substrate is measured at each spot in the array by imaging the entire plate with a commercially available CCD camera. Here, we demonstrate that these 96-well-plate format mini-arrays have performance characteristics that make them suitable for the high-throughput screening of anti-inflammatory compounds.https://www.future-science.com/doi/10.2144/01311dd03
spellingShingle M.D. Moody
S.W. Van Arsdell
K.P. Murphy
S.F. Orencole
C. Burns
Array-Based ELISAs for High-Throughput Analysis of Human Cytokines
BioTechniques
title Array-Based ELISAs for High-Throughput Analysis of Human Cytokines
title_full Array-Based ELISAs for High-Throughput Analysis of Human Cytokines
title_fullStr Array-Based ELISAs for High-Throughput Analysis of Human Cytokines
title_full_unstemmed Array-Based ELISAs for High-Throughput Analysis of Human Cytokines
title_short Array-Based ELISAs for High-Throughput Analysis of Human Cytokines
title_sort array based elisas for high throughput analysis of human cytokines
url https://www.future-science.com/doi/10.2144/01311dd03
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