Analysis of <i>Leishmania chagasi</i> by 2-D Difference Gel Eletrophoresis (2-D DIGE) and Immunoproteomic: Identification of Novel Candidate Antigens for Diagnostic Tests and Vaccine
Míriam M. Costa
Hélida M. Andrade
Daniella C. Bartholomeu
Leandro M. Freitas
Simone F. Pires
Alexander D. Chapeaurouge
Jonas Perales
André T. Ferreira
Mário S. Giusta
Maria N. Melo
Ricardo T. Gazzinelli
10.1021/pr101286y.s001
https://acs.figshare.com/articles/journal_contribution/Analysis_of_i_Leishmania_chagasi_i_by_2_D_Difference_Gel_Eletrophoresis_2_D_DIGE_and_Immunoproteomic_Identification_of_Novel_Candidate_Antigens_for_Diagnostic_Tests_and_Vaccine/2656474
Identification of novel antigens is essential for developing new diagnostic tests and vaccines. We used DIGE to compare protein expression in amastigote and promastigote forms of <i>Leishmania chagasi</i>. Nine hundred amastigote and promastigote spots were visualized. Five amastigote-specific, 25 promastigote-specific, and 10 proteins shared by the two parasite stages were identified. Furthermore, 41 proteins were identified in the Western blot employing 2-DE and sera from infected dogs. From these proteins, 3 and 38 were reactive with IgM and total IgG, respectively. The proteins recognized by total IgG presented different patterns in terms of their recognition by IgG1 and/or IgG2 isotypes. All the proteins selected by Western blot were mapped for B-cell epitopes. One hundred and eighty peptides were submitted to SPOT synthesis and immunoassay. A total of 25 peptides were shown of interest for serodiagnosis to visceral leishmaniasis. In addition, all proteins identified in this study were mapped for T cell epitopes by using the NetCTL software, and candidates for vaccine development were selected. Therefore, a large-scale screening of <i>L. chagasi</i> proteome was performed to identify new B and T cell epitopes with potential use for developing diagnostic tests and vaccines.
2011-05-06 00:00:00
Western blot
IgG
Novel Candidate Antigens
DIGE
T cell epitopes
vaccine
Leishmania chagasi
DE
protein