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Article

Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis

1
Molecular Biology Institute of Paraná, Curitiba, Paraná 81350-010, Brazil
2
Gonçalo Moniz Institute, Oswaldo Cruz Foundation, Salvador, Bahia 40296-710, Brazil
3
Carlos Chagas Institute, Oswaldo Cruz Foundation, Curitiba, Paraná 81350-010, Brazil
4
Integrated Translational Program in Chagas Disease from Fiocruz (Fio-Chagas), Vice Presidency of Research and Biological Collections, Oswaldo Cruz Foundation, Rio de Janeiro, Rio de Janeiro 21040-900, Brazil
*
Author to whom correspondence should be addressed.
Biosensors 2021, 11(8), 289; https://doi.org/10.3390/bios11080289
Received: 3 June 2021 / Revised: 25 July 2021 / Accepted: 26 July 2021 / Published: 22 August 2021
(This article belongs to the Special Issue Biosensing and Bioimaging: Trends and Perspective)
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. View Full-Text
Keywords: Chagas disease; immunoassays; chimeric proteins; stability Chagas disease; immunoassays; chimeric proteins; stability
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MDPI and ACS Style

Celedon, P.A.F.; Leony, L.M.; Oliveira, U.D.; Freitas, N.E.M.; Silva, Â.A.O.; Daltro, R.T.; Santos, E.F.; Krieger, M.A.; Zanchin, N.I.T.; Santos, F.L.N. Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis. Biosensors 2021, 11, 289. https://doi.org/10.3390/bios11080289

AMA Style

Celedon PAF, Leony LM, Oliveira UD, Freitas NEM, Silva ÂAO, Daltro RT, Santos EF, Krieger MA, Zanchin NIT, Santos FLN. Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis. Biosensors. 2021; 11(8):289. https://doi.org/10.3390/bios11080289

Chicago/Turabian Style

Celedon, Paola A.F., Leonardo M. Leony, Ueriton D. Oliveira, Natália E.M. Freitas, Ângelo A.O. Silva, Ramona T. Daltro, Emily F. Santos, Marco A. Krieger, Nilson I.T. Zanchin, and Fred L.N. Santos. 2021. "Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis" Biosensors 11, no. 8: 289. https://doi.org/10.3390/bios11080289

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