Chagas Disease Diagnosis with Trypanosoma cruzi-Exclusive Epitopes in GFP

Author(s)

Durans, Andressa da M. ; Napoleao-Pego, Paloma ; Reis, Flavia C. G. ; Dias, Evandro R. ; Machado, Luciana E. S. F. ; Lechuga, Guilherme C. ; Junqueira, Angela C. V. ; De-Simone, Salvatore G. ; Provance Jr, David W.

• Document type: Journal article
• Source: VACCINES; v. 12, n. 9, p. 17-pg., 2024-09-01.

Abstract

Serological tests are critical tools in the fight against infectious disease. They detect antibodies produced during an adaptive immune response against a pathogen with an immunological reagent, whose antibody binding characteristics define the specificity and sensitivity of the assay. While pathogen proteins have conveniently served as reagents, their performance is limited by the natural grouping of specific and non-specific antibody binding sites, epitopes. An attractive solution is to build synthetic proteins that only contains pathogen-specific epitopes, which could theoretically reach 100% specificity. However, the genesis of de novo proteins remains a challenge. To address the uncertainty of producing a synthetic protein, we have repurposed the beta barrel of fluorescent proteins into a receptacle that can receive several epitope sequences without compromising its ability to be expressed. Here, two versions of a multiepitope protein were built using the receptacle that differ by their grouping of epitopes specific to the parasite Trypanosoma cruzi, the causative agent for Chagas disease. An evaluation of their performance as the capture reagent in ELISAs showed near-complete agreement with recommended diagnostic protocols. The results suggest that a single assay could be developed for the diagnosis of Chagas disease and that this approach could be applied to other diseases. (AU)

• FAPESP's process: 19/02605-3 - Structural and functional characterization of mitochondrial protein phosphatases by redox metabolism and their role in cell biology
• Grantee: Luciana Elena de Souza Fraga Machado
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 20/10168-0 - Structural and functional characterization of mitochondrial protein phosphatases by redox metabolism and their role in cell biology
• Grantee: Luciana Elena de Souza Fraga Machado
• Support Opportunities: Scholarships in Brazil - Young Researchers