Engineering a single-chain antibody against Trypanosoma cruzi metacyclic trypomastigotes to block cell invasion
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Fecha
2019-10
Autores
Kalempa Demeu, Lara Maria
Jahn Soares, Rodrigo
Severo Miranda, Juliana
Pacheco-Lugo, Lisandro A.
Gonçalves Oliveira, Kelin
Cortez Plaza, Cristian Andrés
Billiald, Philippe
Ferreira de Moura, Juliana
Yoshida, Nobuko
Magalhães Alvarenga, Larissa
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Javier Marcelo Di Noia, Institut de recherches cliniques de Montreal, CANADA
Resumen
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.
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Palabras clave
Trypanosoma cruzi, Parasitic diseases, Periplasm, Chagas disease, Protein extraction, Protozoan infections, Sequence databases, Trypomastigotes
