title: mAb CZP-315.D9: an antirecombinant cruzipain monoclonal antibody that specifically labels the reservosomes of Trypanosoma cruzi epimastigotes.
abstract: Reservosomes are large round vesicles located at the posterior end of epimastigote forms of the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. They are the specific end organelles of the endocytosis pathway of T. cruzi, and they play key roles in nutrient uptake and cell differentiation. These lysosome-like organelles accumulate ingested macromolecules and contain large amounts of a major cysteine proteinase (cruzipain or GP57/51 protein). Aim of this study was to produce a monoclonal antibody (mAb) against a recombinant T. cruzi cruzipain (TcCruzipain) that specifically labels the reservosomes. BALB/c mice were immunized with purified recombinant TcCruzipain to obtain the mAb. After fusion of isolated splenocytes with myeloma cells and screening, a mAb was obtained by limiting dilution and characterized by capture ELISA. We report here the production of a kappa-positive monoclonal IgG antibody (mAb CZP-315.D9) that recognizes recombinant TcCruzipain. This mAb binds preferentially to a protein with a molecular weight of about 50 kDa on western blots and specifically labels reservosomes by immunofluorescence and transmission electron microscopy. The monoclonal CZP-315.D9 constitutes a potentially powerful marker for use in studies on the function of reservosomes of T. cruzi.
authors.affiliation: Laboratório de Biologia Celular, Instituto Carlos Chagas/Fiocruz, Rua Professor Algacyr Munhóz Mader 3775, Cidade Industrial, 81350-010 Curitiba, PR, Brazil.
authors.forename: Cassiano Martin
authors.initials: CM
authors.lastname: Batista
authors.affiliation: Laboratório de Biologia Celular, Instituto Carlos Chagas/Fiocruz, Rua Professor Algacyr Munhóz Mader 3775, Cidade Industrial, 81350-010 Curitiba, PR, Brazil.
authors.forename: Lia Carolina Soares
authors.initials: LC
authors.lastname: Medeiros
authors.affiliation: Laboratório de Biologia Celular, Instituto Carlos Chagas/Fiocruz, Rua Professor Algacyr Munhóz Mader 3775, Cidade Industrial, 81350-010 Curitiba, PR, Brazil.
authors.forename: Iriane
authors.initials: I
authors.lastname: Eger
authors.affiliation: Laboratório de Biologia Celular, Instituto Carlos Chagas/Fiocruz, Rua Professor Algacyr Munhóz Mader 3775, Cidade Industrial, 81350-010 Curitiba, PR, Brazil.
authors.forename: Maurilio José
authors.initials: MJ
authors.lastname: Soares
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage* / Q000276:immunology; D002454:Cell Differentiation; D014355:Chagas Disease / Q000469:parasitology / Q000473:pathology*; D003546:Cysteine Endopeptidases / Q000276:immunology / Q000378:metabolism*; D004705:Endocytosis / Q000187:drug effects / Q000276:immunology; D006801:Humans; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D046529:Microscopy, Electron, Transmission; D015388:Organelles / Q000473:pathology / Q000648:ultrastructure; D015800:Protozoan Proteins; D014349:Trypanosoma cruzi / Q000276:immunology / Q000378:metabolism* / Q000472:pathogenicity
chemical_list: D000911:Antibodies, Monoclonal; D015800:Protozoan Proteins; D003546:Cysteine Endopeptidases; C063214:cruzipain