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Purification of Glycosylphosphatidylinositol-Anchored Mucins from Trypanosoma cruzi Trypomastigotes and Synthesis of α-Gal-Containing Neoglycoproteins: Application as Biomarkers for Reliable Diagnosis and Early Assessment of Chemotherapeutic Outcomes of Chagas Disease

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T. cruzi Infection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1955))

Abstract

Chagas disease (ChD), caused by the protozoan parasite Trypanosoma cruzi, affects millions of people worldwide. Chemotherapy is restricted to two drugs, which are partially effective and may cause severe side effects, leading to cessation of treatment in a significant number of patients. Currently, there are no biomarkers to assess therapeutic efficacy of these drugs in the chronic stage. Moreover, no preventive or therapeutic vaccines are available. In this chapter, we describe the purification of Trypanosoma cruzi trypomastigote-derived glycosylphosphatidylinositol (GPI)-anchored mucins (tGPI-mucins) for their use as antigens for the reliable primary or confirmatory diagnosis and as prognostic biomarkers for early assessment of cure following ChD chemotherapy. We also describe, as an example, the synthesis of a potential tGPI-mucin-derived α-Gal-terminating glycan and its coupling to a carrier protein for use as diagnostic and prognostic biomarker in ChD.

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Acknowledgments

This study was funded by the NIH grants 1U01AI129783-01A1 (to ICA, JG, and FT), 1R21AI07961801A1 (to KM) and 1R21AI115451 (to ICA), and UTEP College of Science Multidisciplinary Pilot Projects and Collaborations Grant (to ICA and KM). We would like to thank the UTEP/BBRC Biomolecule Analysis Core Facility (BACF) for access to the LC–MS and MS instruments. All BBRC/UTEP Core Facilities were supported by a grant (2G12MD007592) from the National Institute on Minority Health and Health Disparities (NIMHD). SP was recipient of Dr. Keelung Hong Graduate Research Fellowship and UTEP Dodson Research Grant. EI was the recipient of the Diana Natalicio Doctoral Dissertation Fellowship, UTEP. UOR and EI were fellows of the RISE Research Scholars Program (R25GM069621-11 to Dr. Renato Aguilera, PI). NKH was recipient of Frank B. Cotton Trust Scholarship, from UTEP Graduate School. We are grateful to Dr. Sergio Sosa-Estani, Drugs for Neglected Diseases Initiative (DNDi), for kindly providing travel funds to ON to visit the Dept. of Biological Sciences, University of Texas at El Paso. ICA, FT, JG, MJP, ON, and BAN are members of the NHEPACHA (New Tools for the Diagnosis and Evaluation of Chagas Disease) network.

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Author notes

  1. Susana Portillo, Roger A. Ashmus, and Uriel Ortega-Rodriguez have equally contributed to this chapter.

Authors and Affiliations

  1. Department of Biological Sciences, University of Texas at El Paso, El Paso, TX, USA

    Uriel Ortega-Rodriguez, Susana Portillo, Jerry A. Duran, Eva Iniguez, Brenda G. Zepeda, Janet J. Olivas, Nasim H. Karimi, Rosa A. Maldonado & Igor C. Almeida

  2. Department of Chemistry and Biochemistry, University of Texas at El Paso, El Paso, TX, USA

    Roger A. Ashmus, Nathaniel S. Schocker, Alba L. Montoya & Katja Michael

  3. Barcelona Centre for International Health Research (ISGlobal), Barcelona, Spain

    Luis Izquierdo & Maria-Jesús Pinazo

  4. Facultad de Medicina, Instituto de Medicina Tropical, Universidad Central de Venezuela, Caracas, Venezuela

    Belkisyolé Alarcón de Noya, Oscar Noya & Joaquim Gascon

  5. Faculty of Medicine, Universidad Mayor de San Simón, Cochabamba, Bolivia

    Faustino Torrico

  6. Fundación CEADES, Cochabamba, Bolivia

    Faustino Torrico

  7. Barcelona Institute for Global Health (ISGlobal), Hospital Clínic – Universitat de Barcelona, Barcelona, Spain

    Julio Alonso-Padilla

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  1. Uriel Ortega-Rodriguez
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  2. Susana Portillo
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  3. Roger A. Ashmus
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  20. Igor C. Almeida
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Corresponding author

Correspondence to Igor C. Almeida .

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Editors and Affiliations

  1. INGEBI-CONICET, Buenos Aires, Argentina

    Karina Andrea Gómez

  2. UNSAM-CONICET, IIB-INTECH, San Martin, Buenos Aires, Argentina

    Carlos Andrés Buscaglia

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Ortega-Rodriguez, U. et al. (2019). Purification of Glycosylphosphatidylinositol-Anchored Mucins from Trypanosoma cruzi Trypomastigotes and Synthesis of α-Gal-Containing Neoglycoproteins: Application as Biomarkers for Reliable Diagnosis and Early Assessment of Chemotherapeutic Outcomes of Chagas Disease. In: Gómez, K., Buscaglia, C. (eds) T. cruzi Infection. Methods in Molecular Biology, vol 1955. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9148-8_22

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  • DOI: https://doi.org/10.1007/978-1-4939-9148-8_22

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