Comparative Evaluation of Cytokine Assays

The cytokine assays tested in this study included traditional solid-phase sandwich enzyme-linked immunosorbent assays (ELISA) utilizing colorimetry or luminometry platforms, immunobead-based assays utilizing the fluidics Luminex fluorometry platform, and a newer class of solid-phase microspot assays based on ultra-low-noise charge-coupled device (CCD) cameras represented by the Meso Scale Discovery (MSD) electrochemiluminescence platform. ELISA assays were obtained from Invitrogen BioSource (Carlsbad, CA), R&D Systems (Minneapolis, MN), and Pierce Endogen (Rockford, IL). Luminex assay kits were either purchased from Millipore Upstate (Charlottesville, VA), or were assembled in-house using reagents from R&D Systems. For comparison purposes in this paper, these two assays are labeled as Beadlyte Luminex and in-house Luminex. MSD assay kits were purchased from Meso Scale Discovery (Gaithersburg, MD). A detailed description of immunoassays is provided in Table 1.

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Fichorova R.N., Richardson-Harman N., Alfano M., Belec L., Carbonneil C., Chen S., Cosentino L., Curtis K., Dezzutti C.S., Donoval B., Doncel G.F., Donaghay M., Grivel J.C., Guzman E., Hayes M., Herold B., Hillier S., Lackman-Smith C., Landay A., Margolis L., Mayer K.H., Pasicznyk J.M., Pallansch-Cokonis M., Poli G., Reichelderfer P., Roberts P., Rodriguez I., Saidi H., Sassi R.R., Shattock R, & Cummins, Jr. J.E. (2008). Biological and Technical Variables Affecting Immunoassay Recovery of Cytokines from Human Serum and Simulated Vaginal Fluid: A Multicenter Study. Analytical Chemistry, 80(12), 4741-4751.

Publication 2008

Assay Colorimetry Cytokine Device Enzyme linked immunosorbent assays Fluorometry Immunoassays

Corresponding Organization :

Other organizations : Southern Research Institute, Magee-Womens Research Institute, Vita-Salute San Raffaele University, Brigham and Women's Hospital, National Institutes of Health

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Variable analysis

independent variables

Type of immunoassay platform (ELISA, Luminex, MSD)

dependent variables

Cytokine levels or concentrations

control variables

Reagents used in the different immunoassay platforms (e.g., Invitrogen BioSource, R&D Systems, Pierce Endogen, Millipore Upstate, Meso Scale Discovery)

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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