Purification of Antibodies Against Entamoeba histolytica MIF and Their Use in Analyzing Human and Mouse Samples

  • Laura Farr
  • Koji Watanabe
  • Shannon MoonahEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2080)


Macrophage migration inhibitory factor (MIF) is a proinflammatory and proproliferative cytokine expressed in humans. MIF homologs also exist in many pathogenic protozoans, including Entamoeba, Plasmodium, Toxoplasma, and Leishmania. Production of antibodies against parasite proteins allows for the generation of assays to measure and visualize parasite infection within hosts. In this chapter, we describe how to specifically purify antibodies against Entamoeba histolytica MIF (EhMIF), and subsequently use anti-EhMIF antibodies for ELISA on mouse and human samples and for immunohistochemistry on human tissue. These methods can be applied to any protein for high-quality antibody purification.

Key words

Amebiasis Entamoeba Parasite 


  1. 1.
    Ghosh S et al (2018) Interaction between parasite-encoded JAB1/CSN5 and macrophage migration inhibitory factor proteins attenuates its proinflammatory function. Sci Rep 8(1):10241CrossRefGoogle Scholar
  2. 2.
    Ngobeni R et al (2017) Entamoeba histolytica-encoded homolog of macrophage migration inhibitory factor contributes to mucosal inflammation during amebic colitis. J Infect Dis 215(8):1294–1302CrossRefGoogle Scholar
  3. 3.
    Moonah SN et al (2014) The macrophage migration inhibitory factor homolog of Entamoeba histolytica binds to and immunomodulates host macrophages. Infect Immun 82(9):3523–3530CrossRefGoogle Scholar
  4. 4.
    Shirley D-AT et al (2018) A review of the global burden, new diagnostics, and current therapeutics for amebiasis. Open Forum Infect Dis 5(7):ofy161CrossRefGoogle Scholar
  5. 5.
    Moonah S et al (2014) Erythrocyte lysis and Xenopus laevis oocyte rupture by recombinant Plasmodium falciparum hemolysin III. Eukaryot Cell 13(10):1337–1345CrossRefGoogle Scholar
  6. 6.
    Mori K et al (2013) The C9orf72 GGGGCC repeat is translated into aggregating dipeptide-repeat proteins in FTLD/ALS. Science 339(6125):1335–1338CrossRefGoogle Scholar
  7. 7.
    Annis MG et al (2001) Endoplasmic reticulum localized Bcl-2 prevents apoptosis when redistribution of cytochrome c is a late event. Oncogene 20:1939CrossRefGoogle Scholar
  8. 8.
    Watanabe K et al (2017) Microbiome-mediated neutrophil recruitment via CXCR2 and protection from amebic colitis. PLoS Pathog 13(8):e1006513CrossRefGoogle Scholar
  9. 9.
    Schlaepfer DD et al (1994) Integrin-mediated signal transduction linked to Ras pathway by GRB2 binding to focal adhesion kinase. Nature 372(22):786–791CrossRefGoogle Scholar
  10. 10.
    González-Canto A, et al (2017) Detection of specific antibodies for amoebapore in serum of patients with amoebic liver abscess. Ann Clin Pathol 5(1):1103–1110Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.University of Virginia School of MedicineCharlottesvilleUSA
  2. 2.National Center for Global Health and MedicineShinjukuJapan
  3. 3.Department of MedicineUniversity of Virginia School of MedicineCharlottesvilleUSA

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