Engineering a single-chain antibody against Trypanosoma cruzi metacyclic trypomastigotes to block cell invasion
Autoři:
Lara Maria Kalempa Demeu aff001; Rodrigo Jahn Soares aff001; Juliana Severo Miranda aff002; Lisandro A. Pacheco-Lugo aff001; Kelin Gonçalves Oliveira aff001; Cristian Andrés Cortez Plaza aff004; Philippe Billiald aff006; Juliana Ferreira de Moura aff002; Nobuko Yoshida aff004; Larissa Magalhães Alvarenga aff002; Wanderson Duarte DaRocha aff001
Působiště autorů:
Departamento de Bioquímica e Biologia Molecular, Setor de Ciências Biológicas, Universidade Federal do Paraná,Curitiba, Brasil
aff001; Departamento de Patologia Básica, Setor de Ciências Biológicas, Universidade Federal do Paraná, Curitiba, Brasil
aff002; Universidad Simón Bolívar, Barranquilla, Colombia
aff003; Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brasil
aff004; Centro de Genómica y Bioinformática, Facultad de Ciencias, Universidad Mayor, Santiago, Chile
aff005; Faculte de Pharmacie, Universite Paris-Sud, Paris, France
aff006
Vyšlo v časopise:
PLoS ONE 14(10)
Kategorie:
Research Article
doi:
https://doi.org/10.1371/journal.pone.0223773
Souhrn
Trypanosoma cruzi is a flagellate protozoan pathogen that causes Chagas disease. Currently there is no preventive treatment and the efficiency of the two drugs available is limited to the acute phase. Therefore, there is an unmet need for innovative tools to block transmission in endemic areas. In this study, we engineered a novel recombinant molecule able to adhere to the T. cruzi surface, termed scFv-10D8, that consists of a single-chain variable fragment (scFv) derived from mAb-10D8 that targets gp35/50. The synthetic gene encoding scFv-10D8 was cloned and fused to a 6×His tag and expressed in a prokaryotic expression system. Total periplasmic or 6xHis tag affinity-purified fractions of scFv-10D8 retained the capacity to bind to gp35/50, as shown by Western blot analyses. Pre-incubation of metacyclic trypomastigotes with scFv-10D8 showed a remarkable reduction in cell invasion capacity. Our results suggest that scFv-10D8 can be used in a paratransgenic approach to target parasites in insect vectors, avoiding dissemination of infective forms. Such advances in the development of this functional molecule will surely prompt the improvement of alternative strategies to control Chagas disease by targeting mammalian host stages.
Klíčová slova:
Chagas disease – Parasitic diseases – Protein extraction – Protozoan infections – Sequence databases – Trypanosoma cruzi – Periplasm – Trypomastigotes
Zdroje
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