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Allergy pp 91–114Cite as

Improved Methods for Quantifying Human Chemokine and Cytokine Biomarker Responses: Ultrasensitive ELISA and Meso Scale Electrochemiluminescence Assays

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2020))

Abstract

ELISAs and similar immunoassays are a backbone of biomedical research and clinical practice. Here we review the major factors to consider in the development and application of ultrasensitive ELISAs for analysis of human immune responses in plasma, serum, urine, or tissue culture supernatants. We focus on cytokine and chemokine biomarkers of health and chronic inflammatory diseases including allergy, asthma, autoimmunity, and cardiovascular disease. Detailed protocols for ELISA and Meso Scale Discovery assays (an improved variant of ELISA) are provided for 15 cytokines and 11 chemokines that play immune-regulatory roles in human innate and adaptive immunity. Protocols have been individually optimized to yield ultrasensitive limits of detection and quantification. Major factors enhancing immunoassay sensitivity, precision, and reproducibility, as well as key pitfalls in assay design and execution, are critically reviewed.

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References

  1. HayGlass KT (2012) Complex asthma endotypes, differential chemokine responses and birth cohort studies: solving equations with multiple variables. Clin Exp Allergy 42(11):1546–1548

    Article  CAS  Google Scholar 

  2. HayGlass KT (2011) The quest for predictive immune biomarkers. Clin Exp Allergy 41(12):1656–1659

    Article  CAS  Google Scholar 

  3. Maciejko L, Smalley M, Goldman A (2017) Cancer immunotherapy and personalized medicine: emerging technologies and biomarker-based approaches. J Mol Biomark Diagn 8(5):350

    Article  Google Scholar 

  4. Alexis NE (2014) Biomarker sampling of the airways in asthma. Curr Opin Pulm Med 20(1):46–52

    Article  CAS  Google Scholar 

  5. Tunceroglu H, Shah A, Porhomayon J et al (2013) Biomarkers of lung injury in critical care medicine: past, present, and future. Immunol Investig 42(3):247–261

    Article  CAS  Google Scholar 

  6. Easton R, Balogh ZJ (2014) Peri-operative changes in serum immune markers after trauma: a systematic review. Injury 45(6):934–941

    Article  Google Scholar 

  7. Massaro AN, Wu YW, Bammler TK et al (2018) Plasma biomarkers of brain injury in neonatal hypoxic-ischemic encephalopathy. J Pediatr 194:67–75 e1

    Article  CAS  Google Scholar 

  8. Dhondalay GK, Rael E, Acharya S et al (2018) Food allergy and omics. J Allergy Clin Immunol 141(1):20–29

    Article  CAS  Google Scholar 

  9. Prasad G, McCullough M (2013) Chemokines and cytokines as salivary biomarkers for the early diagnosis of oral cancer. Int J Dent 2013:813756

    Article  Google Scholar 

  10. Rodney T, Osier N, Gill J (2018) Pro- and anti-inflammatory biomarkers and traumatic brain injury outcomes: a review. Cytokine 110:248–256. https://doi.org/10.1016/j.cyto.2018.01.012

    Article  CAS  PubMed  Google Scholar 

  11. Henriksen K, O’Bryant SE, Hampel H et al (2014) The future of blood-based biomarkers for Alzheimer’s disease. Alzheimers Dement 10(1):115–131

    Article  Google Scholar 

  12. Yalow RS, Berson SA (1960) Immunoassay of endogenous plasma insulin in man. J Clin Invest 39:1157–1175

    Article  CAS  Google Scholar 

  13. Imada M, Simons FE, Jay FT et al (1995) Antigen mediated and polyclonal stimulation of human cytokine production elicit qualitatively different patterns of cytokine gene expression. Int Immunol 7(2):229–237

    Article  CAS  Google Scholar 

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Acknowledgments

We thank the many colleagues and collaborators who have provided suggestions which aided in the development of these methodologies. We acknowledge funding from the CIHR (Canadian Institutes for Health Research) and the Canada Research Chairs program.

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Authors and Affiliations

  1. Department of Immunology, University of Manitoba, Winnipeg, MB, Canada

    William P. Stefura, Caroline Graham & Kent T. HayGlass

  2. Department of Community Health and Epidemiology, Health Science Building, University of Saskatchewan, Saskatoon, SK, Canada

    Larisa Lotoski

Authors
  1. William P. Stefura
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  2. Caroline Graham
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  3. Larisa Lotoski
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  4. Kent T. HayGlass
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Corresponding author

Correspondence to Kent T. HayGlass .

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Editors and Affiliations

  1. Core Facilities, St. George’s University of London, London, UK

    Penny Lympany

  2. Department of Occupational and Environmental Medicine, Imperial College London, London, UK

    Meinir G. Jones

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Stefura, W.P., Graham, C., Lotoski, L., HayGlass, K.T. (2019). Improved Methods for Quantifying Human Chemokine and Cytokine Biomarker Responses: Ultrasensitive ELISA and Meso Scale Electrochemiluminescence Assays. In: Lympany, P., Jones, M. (eds) Allergy. Methods in Molecular Biology, vol 2020. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9591-2_7

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  • DOI: https://doi.org/10.1007/978-1-4939-9591-2_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9589-9

  • Online ISBN: 978-1-4939-9591-2

  • eBook Packages: Springer Protocols

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