Simultaneous measurement of multiple ear proteins with multiplex ELISA assays.
Simultaneous measurement of multiple ear proteins with multiplex ELISA assays.
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DOI：
10.1016/j.heares.2010.11.009
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发表时间：
2011-05
期刊：
影响因子：
2.8
通讯作者：
MacArthur, Carol J.
中科院分区：
文献类型：
--
作者：
Trune, Dennis R.;Larrain, Barbara E.;Hausman, Frances A.;Kempton, J. Beth;MacArthur, Carol J.
A recent advancement in enzyme-linked immunosorbent assay (ELISA) technology is the multiplex antibody array that measures multiple proteins simultaneously within a single sample. This allows reduction in sample volume, time, labor, and material costs, while increasing sensitivity over single ELISA. Current multiplex platforms include planar-based systems using microplates or slides, or bead-based suspension assay with microspheres. To determine the applicability of this technology for ear research, we used 3 different multiplex ELISA-based immunoassay arrays from 4 different companies to measure cytokine levels in mouse middle and inner ear tissue lysate extracts 24 hours following transtympanic Haemophilus influenzae inoculation. Middle and inner ear tissue lysates were analyzed using testing services from Quansys BioSciences, Aushon Biosystems SearchLight (both microplate-based), Milliplex MAP Sample (bead-based), and a RayBiotech, Inc. (slide-based) kit. Samples were assayed in duplicate or triplicate. Results were compared to determine their relative sensitivity and reliability for measures of cytokine related to inflammation. The cytokine pg/ml amounts varied among the multiplex assays, so a comparison also was made of the mean fold increase in cytokines from untreated controls. Several cytokines and chemokines were elevated, the extent dependent upon the assay sensitivity. Those most significantly elevated were IL-1α, IL-1β, IL-6, TNFα, VEGF, IL-8/MIP-2. The results of the multiplex systems were compared with single ELISA kits (IL-1β, IL-6) to assess sensitivity over the traditional method. Overall, the Quansys Biosciences and SearchLight arrays showed the greatest sensitivity, both employing the same multiplex methodology of a spotted array within a microplate well with chemiluminescent detection. They also were more sensitive than the traditional single ELISA performed with commercial kits and matched gene expression changes determined by quantitative RT-PCR. The Quansys array showed a limit of detection for ear IL-6 down to 2–4 pg/ml, indicating it is sufficiently sensitive to detect ear proteins present in low concentrations. Thus, the multiplex ELISA procedures appear suitable and reliable for the study of hearing related proteins, providing accurate, quantitative, reproducible results with considerable improvement in sensitivity and economy.
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影响因子：
2.6
作者：
CURTIS, LM;TENCATE, WJF;RAREY, KE
通讯作者：
RAREY, KE
DOI：
10.1016/j.jpba.2004.05.024
发表时间：
2005-01-04
影响因子：
3.4
作者：
Ray, CA;Bowsher, RR;Dean, RA
通讯作者：
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影响因子：
3.4
作者：
Khan, SS;Smith, MS;McCoy, JP
通讯作者：
McCoy, JP
DOI：
10.1007/978-1-59745-447-6_4
发表时间：
2009
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
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作者：
Young HA
通讯作者：
Young HA
DOI：
10.1093/gerona/63.8.879
发表时间：
2008-08
期刊：
The journals of gerontology. Series A, Biological sciences and medical sciences
影响因子：
--
作者：
Leng SX;McElhaney JE;Walston JD;Xie D;Fedarko NS;Kuchel GA
通讯作者：
Kuchel GA