title: Latex-protein complexes from an acute phase recombinant antigen of Toxoplasma gondii for the diagnosis of recently acquired toxoplasmosis.
abstract: The synthesis and characterization of latex-protein complexes (LPC), from the acute phase recombinant antigen P35 (P35Ag) of Toxoplasma gondii and "core-shell" carboxylated or polystyrene (PS) latexes (of different sizes and charge densities) are considered, with the aim of producing immunoagglutination reagents able to detect recently acquired toxoplasmosis. Physical adsorption (PA) and chemical coupling (CC) of P35Ag onto latex particles at different pH were investigated. Greater amounts of adsorbed protein were obtained on PS latexes than on carboxylated latexes, indicating that hydrophobic forces govern the interactions between the protein and the particle surface. In the CC experiments, the highest amount of bound protein was obtained at pH 6, near the isoelectric point of the protein (IP=6.27). At this pH, it decreased both the repulsion between particle surface and protein, and the repulsion between neighboring molecules. The LPC were characterized and the antigenicity of the P35Ag protein coupled on the particles surface was evaluated by Enzyme-Linked ImmunoSorbent Assay (ELISA). Results from ELISA showed that the P35Ag coupled to the latex particles surface was not affected during the particles sensitization by PA and CC and the produced LPC were able to recognize specific anti-P35Ag antibodies present in the acute phase of the disease.
authors.affiliation: INTEC (Universidad Nacional del Litoral and CONICET), Güemes 3450, Santa Fe 3000, Argentina.
authors.forename: Leandro E
authors.initials: LE
authors.lastname: Peretti
authors.affiliation: INTEC (Universidad Nacional del Litoral and CONICET), Güemes 3450, Santa Fe 3000, Argentina.
authors.forename: Verónica D G
authors.initials: VD
authors.lastname: Gonzalez
authors.affiliation: Laboratorio de Tecnología Inmunológica, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe 3000, Argentina.
authors.forename: Iván S
authors.initials: IS
authors.lastname: Marcipar
authors.affiliation: INTEC (Universidad Nacional del Litoral and CONICET), Güemes 3450, Santa Fe 3000, Argentina. Electronic address: lgug@intec.unl.edu.ar.
authors.forename: Luis M
authors.initials: LM
authors.lastname: Gugliotta
mesh_terms: D000210:Acute-Phase Reaction / Q000276:immunology*; D000327:Adsorption; D000953:Antigens, Protozoan / Q000276:immunology*; D004586:Electrophoresis; D004797:Enzyme-Linked Immunosorbent Assay; D006863:Hydrogen-Ion Concentration; D007840:Latex / Q000737:chemistry / Q000276:immunology*; D011137:Polystyrenes / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology*; D014122:Toxoplasma / Q000276:immunology*; D014123:Toxoplasmosis / Q000175:diagnosis* / Q000276:immunology / Q000469:parasitology
chemical_list: D000953:Antigens, Protozoan; D007840:Latex; D011137:Polystyrenes; D011994:Recombinant Proteins
keywords: Acute phase; ELISA; Latex–protein complex; Toxoplasmosis