title: Production and characterization of monoclonal antibodies against the major cysteine proteinase of Trypanosoma cruzi.
abstract: In the present study we describe the production and characterization of a panel of monoclonal antibodies (MoAbs) directed against cruzipain (Crz), the major cysteine proteinase from Trypanosoma cruzi. The five MoAbs, BD6, BF2, CG2, CH8, and DC10 were analysed with respect to affinity and specificity. None of the MoAbs cross-reacted with papain, which has regions of high homology with Crz. Treatment of the antigen with periodate did not affect the binding of the MoAbs, suggesting that they bind to the polypeptide moiety of Crz. CH8 recognized a continuous epitope located at the C-terminal extension of the proteinase that appeared to be highly immunogenic. Although the rest of the MoAbs recognized epitopes located in the catalytic domain, the enzymatic activity of Crz was not impaired by the binding of the MoAbs. Characterization of the antibody-binding sites revealed the presence of at least four separate epitopes.
authors.affiliation: Cátedra de Inmunología, Facultad de Química, Universidad de la República, Montevideo, Uruguay.
authors.forename: G
authors.initials: G
authors.lastname: Gonzalez
authors.forename: A
authors.initials: A
authors.lastname: Orn
authors.forename: J J
authors.initials: JJ
authors.lastname: Cazzulo
authors.forename: K O
authors.initials: KO
authors.lastname: Grönvik
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000913:Antibodies, Protozoan / Q000096:biosynthesis* / Q000276:immunology; D000915:Antibody Affinity; D000918:Antibody Specificity; D003546:Cysteine Endopeptidases / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D051379:Mice; D008807:Mice, Inbred BALB C; D011233:Precipitin Tests; D015800:Protozoan Proteins; D014349:Trypanosoma cruzi / Q000201:enzymology / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D015800:Protozoan Proteins; D003546:Cysteine Endopeptidases; C063214:cruzipain