Banca de DEFESA: ANDRE VINICIUS FERNANDES FERREIRA

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : ANDRE VINICIUS FERNANDES FERREIRA
DATE: 30/07/2021
TIME: 09:00
LOCAL: Remoto via plataforma Google Meet
TITLE:

RATIONAL DESIGN OF ANTIGENIC TARGETS FOR OBTAINING OF ANTIGENS THAT MAY BE USED IN TESTING OF IMMUNODIAGNOSIS OF BOVINE TRYPANOSOMIASIS

KEY WORDS:

antigenic targets, diagnosis, ELISA, Trypanosoma vivax

PAGES: 67
BIG AREA: Multidisciplinar
AREA: Biotecnologia
SUMMARY:

Bovine trypanosomiasis is a disease caused by protozoa Trypanosoma spp., Among them, the species Trypanosoma vivax (T. vivax). This disease has economic relevance in the livestock sector due to its negative impact on beef cattle. In view of the difficulties encountered for a correct and early diagnosis, immunodiagnosis presents itself as an advantage over other methods due to reliability, ease of automation and implementation in the field. Thus, the objective of this work is to search for new T. vivax antigenic targets, using bioinformatics tools to be used in immunodiagnostic tests for bovine trypanosomiasis. For this, a survey of the main antigenic proteins of T. vivax was carried out in articles and patents. Then, the in silico analysis of the proteins was performed, using bioinformatics programs to select epitopes with antigenic potential. Subsequently, the rational construction of a protein (rTV1) and a synthetic peptide (TV1P) was  performed. Through a partnership with FIOCRUZ-MG, two proteins also candidates for the diagnosis of the disease (M1 and M2) were selected to be produced through heterologous expression, to later be used in biological assays. Proteins TV1, M1 and M2 were used in heterologous expression using Escherichia coli BL21 (λDE3) pLysS and pLysE cells using different expression protocols. Among the protocols used, the expression was successful only with the rTV1 protein, which was purified and used in a preliminary ELISA immunoassay to verify its reactivity towards T. vivax infected sera. The trial suggests that rTV1 has the ability to discriminate between healthy and infected animals. The TV1P peptide was synthesized by chemical synthesis and was used in ELISA assays to evaluate its reactivity towards sera infected with T. vivax. The synthetic peptide was not capable of immunodetection, even though it was evaluated under several conditions. These preliminary data show the feasibility of further research on the rTV1 protein, since it has the potential to be used in diagnostic kits. For the TV1P peptide, further studies are needed in order to evaluate its potential for the diagnosis of T. vivax.

BANKING MEMBERS:
Presidente - 1367304 - ALEXSANDRO SOBREIRA GALDINO
Externo à Instituição - FLORA SATIKO KANO
Externo à Instituição - MARCIO SOBREIRA SILVA ARAUJO - FIOCRUZ
Externa à Instituição - SORAYA DOS SANTOS PEREIRA - FIOCRUZ