Multiplex bead array assays for detection of soluble cytokines: Comparisons of sensitivity and quantitative values among kits from multiple manufacturers

Khan, Sameena S.; Smith, Meghan S.; Reda, Debra; Suffredini, Anthony F.; McCoy, J. Philip

doi:10.1002/cyto.b.20021

Cited by 298 publications

(189 citation statements)

References 18 publications

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“…Indeed, we confirmed that the absolute values of the proteins detected were different between the two tested technologies. Such discordance could be due to differences in the capture and/or detection antibodies and to a differential sensitivity to matrix interference (Dupuy et al, 2013;Khan et al, 2004). However, among the cytokines for which the quantification was consistent, we observed strong inter-platform correlations for IL-8 and VEGF (Table 5), in accordance with previous findings (Chowdhury et al, 2009;Dabitao et al, 2011;Huttunen et al, 2014).…”

Section: Discussionsupporting

confidence: 89%

“…It has already been stressed out that the comparison of absolute cytokine concentration should be attempted only if used reagents were obtained from the same supplier (Khan et al, 2004), although even singleplex assays compared to multiplex assays from the same company can yield significantly different results (de Koning et al, 2012;Huttunen et al, 2014). Indeed, we confirmed that the absolute values of the proteins detected were different between the two tested technologies.…”

Section: Discussionsupporting

confidence: 61%

See 1 more Smart Citation

How to: Measuring blood cytokines in biological psychiatry using commercially available multiplex immunoassays

Belzeaux

Lefebvre

Lazzari

et al. 2017

Psychoneuroendocrinology

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Cytokines produced by both immune and non-immune cells are likely to play roles in the development and/or progression of psychiatric disorders. Indeed, many investigators have compared the blood cytokine levels in psychiatric patients with those of healthy controls or monitored their levels in patients during disease progression to identify biomarkers. Nevertheless, very few studies have confirmed that such cytokines remain stable in healthy individuals through periods of weeks and months. This is an important issue to consider before using blood cytokine levels as biomarkers of disease traits, disease state, or treatment response. Although multiplex assay technology represents an advance in identifying biomarkers because it allows simultaneous examination of large panels of analytes from a small volume of sample, it is necessary to verify whether these assays yield enough sensitivity and reproducibility when applied to the blood from neuropsychiatric patients. Therefore, we compared two multiplex immunoassays, the bead-based Luminex® (Bio-Rad) and the electro-chemiluminescence-based V-plex® (MesoScaleDiscovery), for the detection and quantification of 31 cytokines, chemokines and growth factors in both the sera and plasma of patients with major depressive episodes (MDE) and age-and sex-matched healthy control subjects during a 30-week period. Although both platforms exhibited low coefficients of variability (CV) between the duplicates in the calibration curves, the linearity was better in general for the V-PLEX® platform. However, neither platform was able to detect the absolute values for all of the tested analytes. Among the 16 analytes that were detected by both assays, the intra-assay reproducibility was in general better with the V-PLEX® platform. Although it is not a general rule that the results from sera and plasma will be correlated, consistent results were more frequent with the V-PLEX® platform. Furthermore, the V-PLEX® results were more consistent with the gold standard ELISA simplex assay for IL-6 in both sera and plasma. The intra-individual variability of the measurements, among the sera and plasma for the 4 samples harvested from each healthy individual, was low for Eotaxin, G-CSF, IL-4, IL-7, IL-9, IL-12p40, IL-12p70, IL-15, MIP-1β, PDGF-BB, TNF, TNF-β and VEGF, but intermediate or high for IFN-γ, IL-6, IL-8, IL-10, and IP10. Together, these data suggest that extreme caution is needed in translating the results of multiplex cytokine profiling into biomarker discovery in psychiatry. Dear Editor,We thank you and the Editor in chief, Robert Dantzer, for reaching the conclusion to ultimately publish our manuscript. Following your suggestion, we changed the tiltle to: "How to: Measuring blood cytokines in biological psychiatry using commercially available multiplex immunoassays".We were concerned by the persistent negative comments of Reviewer#3, stressing out that our comparison between the two platforms we have worked with suffers a number of theoretical and technical limitations because...

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Section: Discussionsupporting

confidence: 89%

Section: Discussionsupporting

confidence: 61%

How to: Measuring blood cytokines in biological psychiatry using commercially available multiplex immunoassays

Belzeaux

Lefebvre

Lazzari

et al. 2017

Psychoneuroendocrinology

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show abstract

“…With this in mind, any study that involves sequential monitoring of patients, or other samples, should be performed using only a single technique, one platform, and one commercial vendor for all samples. As illustrated by Khan et al, although multiplex assays from different vendors will show similar trends in cytokine levels, the absolute levels of cytokines measured will vary (22). Therefore, even MBAA assays from different vendors should not be considered interchangeable.…”

Section: Discussionmentioning

confidence: 99%

“…Although commercially available kits have been extensively characterized and qualified by their respective manufacturers, there are a limited number of peer-reviewed reports of direct comparisons between multiplex bead arrays and ELISA kits, and all of these have been published within the last decade (3,5,(21)(22)(23)(24)(25)(26)(27)(28). It is difficult to compare the data from the different studies, as various investigators used different methods of comparison between MBAA and ELISAs, and reports varied considerably in the amount of methodological detail provided.…”

Section: Comparison Of Multiplex Bead Arrays Vs Elisasmentioning

confidence: 99%

Multiplex bead array assays: Performance evaluation and comparison of sensitivity to ELISA☆

Elshal

McCoy

2006

Methods

493

402

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The measurement of soluble cytokines and other analytes in serum and plasma is becoming increasingly important in the study and management of many diseases. As a result, there is a growing demand for rapid, precise, and cost-effective measurement of such analytes in both clinical and research laboratories. Multiplex bead array assays provide quantitative measurement of large numbers of analytes using an automated 96-well plate format. ELISAs (enzyme linked immunosorbent assay) have long been the standard for quantitative analysis of cytokines and other biomarkers, but are not well suited for high throughput multiplex analyses. However, prior to replacement of ELISA assays with multiplex bead array assays, there is a need to know how comparable these two methods are for quantitative analyses. A number of published studies have compared these two methods and it is apparent that certain elements of these assays, such as the clones of monoclonal antibodies used for detection and reporting, are pivotal in obtaining similar results from both assays. By careful consideration of these variables, it should be possible to utilize multiplex bead array assays in lieu of ELISAs for studies requiring high throughput analysis of numerous analytes.

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“…Culture supernatants were harvested after 72 hours and stored at Ϫ80°C until further analyzed. Extracellular cytokines and chemokines were measured using a multiplex immunoassay (Luminex, Austin, TX) as previously described (36)(37)(38). In summary, the antibodycoated microspheres were incubated for 60 minutes with standards or culture medium (25 l) in 96-well, 1.2-m filter plates (Millipore, Amsterdam, The Netherlands).…”

Section: Methodsmentioning

confidence: 99%

PAN–DR‐Binding Hsp60 self epitopes induce an interleukin‐10–mediated immune response in rheumatoid arthritis

Jong¹,

Lafeber²,

Jager³

et al. 2009

Arthritis & Rheumatism

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Objective. Human Hsp60 is expressed in the joints of patients with rheumatoid arthritis (RA) and can elicit a regulatory T cell response in the peripheral blood and synovial fluid. However, Hsp60 can also trigger strong proinflammatory pathways. Thus, to understand the nature of these Hsp60-directed responses in RA, it is necessary to study such responses at the molecular, epitope-specific level. This study was undertaken to characterize the disease specificity and function of pan-DR-binding Hsp60-derived epitopes as possible modulators of autoimmune inflammation in RA.Methods. Lymphocyte proliferation assays (using 3 H-thymidine incorporation and carboxyfluorescein diacetate succinimidyl ester [CFSE] staining) and measurement of cytokine production (using multiplex immunoassay and intracellular staining) were performed after in vitro activation of peripheral blood mononuclear cells from patients with RA, compared with healthy controls. Results. A disease (RA)-specific immune recognition, characterized by T cell proliferation as well as increased production of tumor necrosis factor ␣ (TNF␣), interleukin-1␤ (IL-1␤), and IL-10, was found for 3 of the 8 selected peptides in patients with RA as compared with healthy controls (P < 0.05). Intracellular cytokine staining and CFSE labeling showed that CD4؉ T cells were the subset primarily responsible for both the T cell proliferation and the cytokine production in RA. Interestingly, the human peptides had a remarkably different phenotype, with a 5-10-fold higher IL-10:TNF␣ ratio, compared with that of the microbial peptides.Conclusion. These results suggest a diseasespecific immune-modulatory role of epitope-specific T cells in the inflammatory processes of RA. Therefore, these pan-DR-binding epitopes could be used as a tool to study the autoreactive T cell response in RA and might be suitable candidates for use in immunotherapy.

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Multiplex bead array assays for detection of soluble cytokines: Comparisons of sensitivity and quantitative values among kits from multiple manufacturers

Cited by 298 publications

References 18 publications

How to: Measuring blood cytokines in biological psychiatry using commercially available multiplex immunoassays

How to: Measuring blood cytokines in biological psychiatry using commercially available multiplex immunoassays

Multiplex bead array assays: Performance evaluation and comparison of sensitivity to ELISA☆

PAN–DR‐Binding Hsp60 self epitopes induce an interleukin‐10–mediated immune response in rheumatoid arthritis

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