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https://www.arca.fiocruz.br/handle/icict/49115
STABILITY ASSESSMENT OF FOUR CHIMERIC PROTEINS FOR HUMAN CHAGAS DISEASE IMMUNODIAGNOSIS
Autor(es)
Afiliação
Molecular Biology Institute of Paraná. Curitiba, PR, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.
Molecular Biology Institute of Paraná. Curitiba, PR, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.
Molecular Biology Institute of Paraná. Curitiba, PR, Brazil / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR0, Brasil / Fundação Oswaldo Cruz. Fio-Chagas. Integrated Translational Program in Chagas Disease from Fiocruz. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR0, Brasil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Fio-Chagas. Integrated Translational Program in Chagas Disease from Fiocruz. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.
Molecular Biology Institute of Paraná. Curitiba, PR, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.
Molecular Biology Institute of Paraná. Curitiba, PR, Brazil / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR0, Brasil / Fundação Oswaldo Cruz. Fio-Chagas. Integrated Translational Program in Chagas Disease from Fiocruz. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR0, Brasil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Fio-Chagas. Integrated Translational Program in Chagas Disease from Fiocruz. Rio de Janeiro, RJ, Brasil.
Resumo em Inglês
The performance of an immunoassay relies on antigen-antibody interaction; hence, antigen
chemical stability and structural integrity are paramount for an efficient assay. We conducted a
functional, thermostability and long-term stability analysis of different chimeric antigens (IBMP),
in order to assess effects of adverse conditions on four antigens employed in ELISA to diagnose
Chagas disease. ELISA-based immunoassays have served as a model for biosensors development,
as both assess molecular interactions. To evaluate thermostability, samples were heated and cooled
to verify heat-induced denaturation reversibility. In relation to storage stability, the antigens were
analyzed at 25 C at different moments. Long-term stability tests were performed using eight sets
of microplates sensitized. Antigens were structurally analyzed through circular dichroism (CD),
dynamic light scattering, SDS-PAGE, and functionally evaluated by ELISA. Data suggest that IBMP
antigens are stable, over adverse conditions and for over a year. Daily analysis revealed minor
changes in the molecular structure. Functionally, IBMP-8.2 and IBMP-8.3 antigens showed reactivity
towards anti-T. cruzi antibodies, even after 72 h at 25 C. Long-term stability tests showed that all
antigens were comparable to the control group and all antigens demonstrated stability for one year.
Data suggest that the antigens maintained their function and structural characteristics even in adverse
conditions, making them a sturdy and reliable candidate to be employed in future in vitro diagnostic
tests applicable to different models of POC devices, such as modern biosensors in development
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