Multiplexed analysis of biomarkers related to obesity and the metabolic syndrome in human plasma, using the Luminex-100 system

Mine Y. Liu, Antonios M. Xydakis, Ron C. Hoogeveen, Peter H. Jones, E. O.Brian Smith, Kathleen W. Nelson, Christie M. Ballantyne

Research output: Contribution to journalArticle

138 Citations (Scopus)

Abstract

Background: The complex pathology of disease has sparked the development of novel protein expression profiling techniques that require validation in clinical settings. This study focuses on multiplexed analyses of adipocytokines and biomarkers linked to the metabolic syndrome, diabetes, and cardiovascular disease. Methods: Multiplexed immunoassays using fluorescent microspheres and the Luminex-100 system were performed on plasma from 80 obese patients (40 with the metabolic syndrome) before and after 6-8 weeks of diet-induced weight loss. Leptin, insulin, C-peptide, monocyte chemoattractant protein-1 (MCP-1), eotaxin, interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and IL-6 concentrations measured with multiplex panels from 3 different manufacturers were compared with results from commercial ELISAs. Detection limits and between- and within-run imprecision were determined for each analyte. Bland-Altman analysis was used to determine agreement between multiplexed immunoassays and ELISAs. Results: Correlation between the Luminex multiplexed assays and ELISAs was good for leptin (Linco), insulin (Linco), MCP-1 (Biosource and Upstate), and eotaxin (Biosource) with correlation coefficients of 0.711-0.895; fair for eotaxin (Upstate) and C-peptide (Linco) with correlation coefficients of 0.496-0.582; and poor for TNF-α, IL-8, and IL-6 (Linco, Biosource, Upstate, and R&D) with correlation coefficients of -0.107 to 0.318. Within- and between-run imprecision values for the multiplex method were generally <15%. Relative changes in plasma leptin and insulin concentrations after diet-induced weight loss were similar whether assessed by multiplex assay or ELISA. Conclusion: Although this technology appears useful in clinical research studies, low assay sensitivity and poor correlations with conventional ELISA methods for some analytes with very low plasma concentrations should be considered when using the Luminex platform in clinical studies.

Original languageEnglish
Pages (from-to)1102-1109
Number of pages8
JournalClinical Chemistry
Volume51
Issue number7
DOIs
Publication statusPublished - 2005 Dec 1

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Plasma (human)
Biomarkers
Leptin
Assays
Obesity
Chemokine CCL2
C-Peptide
Enzyme-Linked Immunosorbent Assay
Insulin
Nutrition
Interleukin-8
Plasmas
Interleukin-6
Reducing Diet
Tumor Necrosis Factor-alpha
Adipokines
Immunoassay
Pathology
Medical problems
Microspheres

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Liu, M. Y., Xydakis, A. M., Hoogeveen, R. C., Jones, P. H., Smith, E. O. B., Nelson, K. W., & Ballantyne, C. M. (2005). Multiplexed analysis of biomarkers related to obesity and the metabolic syndrome in human plasma, using the Luminex-100 system. Clinical Chemistry, 51(7), 1102-1109. https://doi.org/10.1373/clinchem.2004.047084
Liu, Mine Y. ; Xydakis, Antonios M. ; Hoogeveen, Ron C. ; Jones, Peter H. ; Smith, E. O.Brian ; Nelson, Kathleen W. ; Ballantyne, Christie M. / Multiplexed analysis of biomarkers related to obesity and the metabolic syndrome in human plasma, using the Luminex-100 system. In: Clinical Chemistry. 2005 ; Vol. 51, No. 7. pp. 1102-1109.
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Multiplexed analysis of biomarkers related to obesity and the metabolic syndrome in human plasma, using the Luminex-100 system. / Liu, Mine Y.; Xydakis, Antonios M.; Hoogeveen, Ron C.; Jones, Peter H.; Smith, E. O.Brian; Nelson, Kathleen W.; Ballantyne, Christie M.

In: Clinical Chemistry, Vol. 51, No. 7, 01.12.2005, p. 1102-1109.

Research output: Contribution to journalArticle

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N2 - Background: The complex pathology of disease has sparked the development of novel protein expression profiling techniques that require validation in clinical settings. This study focuses on multiplexed analyses of adipocytokines and biomarkers linked to the metabolic syndrome, diabetes, and cardiovascular disease. Methods: Multiplexed immunoassays using fluorescent microspheres and the Luminex-100 system were performed on plasma from 80 obese patients (40 with the metabolic syndrome) before and after 6-8 weeks of diet-induced weight loss. Leptin, insulin, C-peptide, monocyte chemoattractant protein-1 (MCP-1), eotaxin, interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and IL-6 concentrations measured with multiplex panels from 3 different manufacturers were compared with results from commercial ELISAs. Detection limits and between- and within-run imprecision were determined for each analyte. Bland-Altman analysis was used to determine agreement between multiplexed immunoassays and ELISAs. Results: Correlation between the Luminex multiplexed assays and ELISAs was good for leptin (Linco), insulin (Linco), MCP-1 (Biosource and Upstate), and eotaxin (Biosource) with correlation coefficients of 0.711-0.895; fair for eotaxin (Upstate) and C-peptide (Linco) with correlation coefficients of 0.496-0.582; and poor for TNF-α, IL-8, and IL-6 (Linco, Biosource, Upstate, and R&D) with correlation coefficients of -0.107 to 0.318. Within- and between-run imprecision values for the multiplex method were generally <15%. Relative changes in plasma leptin and insulin concentrations after diet-induced weight loss were similar whether assessed by multiplex assay or ELISA. Conclusion: Although this technology appears useful in clinical research studies, low assay sensitivity and poor correlations with conventional ELISA methods for some analytes with very low plasma concentrations should be considered when using the Luminex platform in clinical studies.

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