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Profiling of Cytokine and Chemokine Responses Using Multiplex Bead Array Technology

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Immunoproteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1061))

Abstract

Multiplex bead array technology expands upon the principles of the enzyme-linked immunosorbent assay by allowing the simultaneous quantification of a large number of cytokines and chemokines within a single sample. This allows for the researcher more freedom to investigate complex immune responses both in vivo and in vitro. Here we describe the detailed assay protocol and technical tips for simultaneous quantification of multiple cytokines and chemokines in mouse biological fluids such as sera, bronchoalveolar lavage fluid, tissue homogenate supernatant, and tissue culture supernatant, using a multiplex bead array assay.

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References

  1. Khan SS, Smith MS, Reda D, Suffredini AF, McCoy JP Jr (2004) Multiplex bead array assays for detection of soluble cytokines: comparisons of sensitivity and quantitative values among kits from multiple manufacturers. Clin Cytom 61B:35–39

    Article  CAS  Google Scholar 

  2. Elshal MF, McCoy JP Jr (2006) Multiplex bead array assays: performance evaluation and comparison of sensitivity to ELISA. Methods 38:317–323

    Article  PubMed  CAS  Google Scholar 

  3. Fulton RJ, McDade RL, Smith PL, Kienker LJ, Kettman JR Jr (1997) Advanced multiplexed analysis with the FlowMetrix™ system. Clin Chem 43:1749–1756

    PubMed  CAS  Google Scholar 

  4. Keij JF, Steinkamp JA (1998) Flow cytometric characterization and classification of multiple dual-color fluorescent microspheres using fluorescence lifetime. Cytometry 33:318–323

    Article  PubMed  CAS  Google Scholar 

  5. Kettman J, Davies T, Chandler D, Oliver K, Fulton R (1998) Classification and properties of 64 multiplexed microsphere sets. Cytometry 33:234–243

    Article  PubMed  CAS  Google Scholar 

  6. Oliver KG, Kettman JR, Fulton RJ (1998) Multiplexed analysis of human cytokines by use of the FlowMetrix system. Clin Chem 44:2057–2060

    PubMed  CAS  Google Scholar 

  7. Bellisario R, Colinas RJ, Pass KA (2000) Simultaneous measurement of thyroxine and thryotropin from newborn dried blood spot specimens using a multiplexed fluorescent microsphere immunoassay. Clin Chem 46:1422–1424

    PubMed  CAS  Google Scholar 

  8. Taylor JD, Briley D, Nguyen Q, Long K, Iannone MA, Li M-S et al (2001) Flow cytometric platform for high-throughput single nucleotide polymorphism analysis. Biotechniques 30:661–669

    PubMed  CAS  Google Scholar 

  9. de Jager W, te Velthuis H, Prakken BJ, Kuis W, Rijkers GT (2003) Simultaneous detection of 15 human cytokines in a single sample of stimulated peripheral blood mononuclear cells. Clin Diagn Lab Immunol 10:133–139

    PubMed  Google Scholar 

  10. de Jagger W, Prakken BJ, Bijlsma JW, Kuis W, Rijikers GT (2005) Improved multiplex immunoassay performance in human plasma and synovial fluid following removal of interfering heterophilic antibodies. J Immunol Methods 300:124–135

    Article  Google Scholar 

  11. DuPont NC, Wang KH, Wadhwa PD, Culhane JF, Nelson EL (2005) Validation and comparison of Luminex multiplex cytokine analysis kits with ELISA: determinations of a panel of nine cytokines in clinical sample culture supernatants. J Reprod Immunol 66:175–191

    Article  PubMed  CAS  Google Scholar 

  12. Codorean E, Nichita C, Albulescu L, Raducan E, Popescu ID, Lonita AC et al (2010) Correlation of xMAP and ELISA cytokine profiles; development and validation for immunotoxicoligical studies in vitro. Roum Arch Microbiol Immunol 69:13–19

    PubMed  CAS  Google Scholar 

  13. Richens JL, Urbanowiscz RA, Metcalf R, Corne J, O’Shea P, Fairclough L (2010) Quantitative validation and comparison of multiplex cytokine kits. J Biomol Screen 15:562–1572

    Article  PubMed  CAS  Google Scholar 

  14. Searchable bibliography at Luminex website http://www.luminexcorp.com/bibliography

  15. Qiu H, KuoLee R, Harris G, Chen W (2009) High susceptibility to respiratory Acinetobacter baumannii infection in A/J mice is associated with a delay in early pulmonary recruitment of neutrophils. Microbes Infect 11:946–955

    Article  PubMed  CAS  Google Scholar 

  16. Liu M, Xydakis A, Hoogeveen R, Jones P, Smith E, Nelson K et al (2005) Multiplexed analysis of biomarkers related to obesity and the metabolic syndrome in human plasma, using the Luminex-100 system. Clin Chem 51:1102–1109

    Article  PubMed  CAS  Google Scholar 

  17. Landlinger C, Preuner S, Willinger B, Haberpursch B, Racil Z, Mayer J et al (2009) Species-specific identification of a wide range of clinically relevant fungal pathogens by use of Luminex xMAP technology. J Clin Microbiol 47:1063–1073

    Article  PubMed  CAS  Google Scholar 

  18. Joslin E, Opresko L, Wells A, Wiley H, Lauffenburger D (2007) EGF-receptor-mediated mammary epithelial cell migration is driven by sustained ERK signaling from autocrine stimulation. J Cell Sci 120:3688–3699

    Article  PubMed  CAS  Google Scholar 

  19. Scholler N, Crawford M, Sato A, Dreschner C, O'Briant K, Kiviat N et al (2006) Bead-based ELISA for validation of ovarian cancer early detection markers. Clin Cancer Res 12:2117–2124

    Article  PubMed  CAS  Google Scholar 

  20. Vignali DAA (2000) Multiplexed particle based flow cytometric assays. J Immunol Methods 243:243–255

    Article  PubMed  CAS  Google Scholar 

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Harris, G., Chen, W. (2013). Profiling of Cytokine and Chemokine Responses Using Multiplex Bead Array Technology. In: Fulton, K., Twine, S. (eds) Immunoproteomics. Methods in Molecular Biology, vol 1061. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-589-7_16

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  • DOI: https://doi.org/10.1007/978-1-62703-589-7_16

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-588-0

  • Online ISBN: 978-1-62703-589-7

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