Synthesis and Characterization of Latex-protein Complexes from Different Antigens of Toxoplasma gondii for Immunoagglutination Assays

Authors
Peretti, Leandro Ezequiel; Gonzalez, Veronica Doris Guadalupe; Costa, Juan Gabriel; Marcipar, Ivan Sergio; Gugliotta, Luis Marcelino
Publication Year
2016
Language
English
Format
article
Status
Published version
Description
The acute phase recombinant protein of Toxoplasma gondii P22Ag was expressed and purified and thehomogenate of the parasite was obtained from an infected mouse. These antigens were used to producelatex-protein complexes (LPC) through physical adsorption and chemical coupling onto different latexes,with the aim of producing immunoagglutination (IA) reagents able to detect recently acquiredtoxoplasmosis. Polystyrene and ?core-shell? latexes where employed, exhibiting varied particle size,functionality (carboxyl or epoxy), and charge density. In sensitization experiments for producing LPC, therecombinant protein showed better coupling efficiency onto the particles surface than the homogenateand this could be explained by the complex mixture of the homogenate, which includes a large numberof proteins of different molecular mass, isoelectric points, and hydrophobicity. The synthesized LPC wereemployed in IA assays. To this effect, the agglutination reaction was followed by measuring the changesin the optical absorbance by turbidimetry. Experiments against control sera were performed to evaluatethe performance of various LPC and it was observed that the IA test based on P22Ag and thecarboxylated latex of 350 nm of particle diameter allowed a good discrimination between acute sera andchronic/negative ones. The proposed test is cheap, rapid, and easy to implement.
Fil: Peretti, Leandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina
Fil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina
Fil: Costa, Juan Gabriel. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina
Subject
Acute phase recombinant protein
Immunoagglutination
Latex protein complex
Toxoplasmosis
Compuestos
Ingeniería de los Materiales
INGENIERÍAS Y TECNOLOGÍAS
Access level
Restricted access
License
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repository
CONICET Digital (CONICET)
Institution
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identifier
oai:ri.conicet.gov.ar:11336/24185