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

Methods Mol Biol. 2019:1955:287-308. doi: 10.1007/978-1-4939-9148-8_22.

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.

Keywords: Anti-α-Gal antibodies; Biomarkers; Carbohydrate synthesis; Chagas disease; Chemiluminescent enzyme-linked immunosorbent assay; Chemotherapy; Diagnosis; Mucins; Neoglycoproteins; Trypanosoma cruzi.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Chagas Disease / diagnosis*
  • Enzyme-Linked Immunosorbent Assay / methods
  • GPI-Linked Proteins / chemistry
  • GPI-Linked Proteins / isolation & purification*
  • Glycoproteins / chemical synthesis
  • Glycoproteins / chemistry*
  • Humans
  • Macaca mulatta
  • Models, Molecular
  • Mucins / chemistry
  • Mucins / isolation & purification*
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / isolation & purification*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
  • Trypanosoma cruzi / chemistry*

Substances

  • GPI-Linked Proteins
  • Glycoproteins
  • Mucins
  • Protozoan Proteins