Abstract
Immunoaffinity capillary electrophoresis (ICE) is a powerful tool used to detect and quantify target proteins of interest in complex biological fluids. The target analyte is captured and bound to antibodies immobilized onto the wall of a capillary, labeled in situ with a fluorescent dye, eluted and detected online using laser-induced fluorescence following electrophoretic separation. Here, we illustrate how to construct an immunoaffinity capillary and utilize it to run ICE in order to capture and quantify target cytokines and chemokines from a clinical sample.
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Bergquist J et al (1997) Measurements of catecholamine-mediated apoptosis of immunocompetent cells by capillary electrophoresis. Electrophoresis 18:1760–1766
Zhang H, Jin W (2004) Determination of different forms of human interferon-gamma in single natural killer cells by capillary electrophoresis with on-capillary immunoreaction and laser-induced fluorescence detection. Electrophoresis 25:1090–1095
Liang A et al (2008) Further characterization of the binding of heparin to granulocyte colony-stimulating factor: importance of sulfate groups. Electrophoresis 29:1286–1290
Phillips TM, Dickens BF (1998) Analysis of recombinant cytokines in human body fluids by immunoaffinity capillary electrophoresis. Electrophoresis 19:2991–2996
Guzman NA (2000) Determination of immunoreactive gonadotropin-releasing hormone in serum and urine by on-line immunoaffinity capillary electrophoresis coupled to mass spectrometry. J Chromatogr B 749:197–213
Guzman NA, Phillips TM (2005) Immunoaffinity CE for proteomics studies. Anal Chem 77:61A–67A
Phillips TM, Smith P (2003) Analysis of intracellular regulatory proteins by immunoaffinity capillary electrophoresis coupled with laser-induced fluorescence detection. Biomed Chromatogr 17:182–187
Benavente F et al (2007) Determination of human erythropoietin by on-line immunoaffinity capillary electrophoresis: a preliminary report. Anal Bioanal Chem 387:2633–2639
Kuchen S et al (2007) Essential role of IL-21 in B cell activation, expansion, and plasma cell generation during CD4+ T cell-B cell collaboration. J Immunol 179:5886–5896
Kalish H, Phillips TM (2010) Analysis of neurotrophins in human serum by immunoaffinity capillary electrophoresis (ICE) following traumatic head injury. J Chromatogr B 878:194–200
Shah JP et al (2008) Biochemicals associated with pain and inflammation are elevated in sites near to and remote from active myofascial trigger points. Arch Phys Med Rehabil 89: 16–23
Phillips TM (2001) Analysis of single-cell cultures by immunoaffinity capillary electrophoresis with laser-induced fluorescence detection. Luminescence 16:145–152
Summy-Long JY et al (2006) Response of interleukin-1beta in the magnocellular system to salt-loading. J Neuroendocrinol 18:926–937
Guzman NA, Blanc T, Phillips TM (2008) Immunoaffinity capillary electrophoresis as a powerful strategy for the quantification of low-abundance biomarkers, drugs, and metabolites in biological matrices. Electrophoresis 29:3259–3278
Phillips TM, Dickens BF (2000) Affinity and immunoaffinity purification techniques. Eaton publishing, Natick, Massachusetts, p 43
Phillips TM, Dickens BF (2000) Affinity and immunoaffinity purification techniques. Eaton publishing, Natick, Massachusetts, p 129
Phillips TM, Dickens BF (2000) Ligand attachment to supports. In: Affinity and immunoaffinity purification techniques. Eaton publishing, Natick, Massachusetts, p 103
Phillips TM, Dickens BF (2000) Affinity and immunoaffinity purification techniques. Eaton publishing, Natick, Massachusetts, p 297
Phillips TM, Kennedy LM, De Fabo EC (1997) Microdialysis-immunoaffinity capillary electrophoresis studies on neuropeptide-induced lymphocyte secretion. J Chromatogr B 697:101–109
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Mendonca, M., Kalish, H. (2013). Cytokine Analysis by Immunoaffinity Capillary Electrophoresis. In: Phillips, T., Kalish, H. (eds) Clinical Applications of Capillary Electrophoresis. Methods in Molecular Biology, vol 919. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-029-8_23
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DOI: https://doi.org/10.1007/978-1-62703-029-8_23
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-028-1
Online ISBN: 978-1-62703-029-8
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