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Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis
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 as...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8391164/ https://www.ncbi.nlm.nih.gov/pubmed/34436091 http://dx.doi.org/10.3390/bios11080289 |
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author | Celedon, Paola Alejandra Fiorani Leony, Leonardo Maia Oliveira, Ueriton Dias Freitas, Natália Erdens Maron Silva, Ângelo Antônio Oliveira Daltro, Ramona Tavares Santos, Emily Ferreira Krieger, Marco Aurélio Zanchin, Nilson Ivo Tonin Santos, Fred Luciano Neves |
author_facet | Celedon, Paola Alejandra Fiorani Leony, Leonardo Maia Oliveira, Ueriton Dias Freitas, Natália Erdens Maron Silva, Ângelo Antônio Oliveira Daltro, Ramona Tavares Santos, Emily Ferreira Krieger, Marco Aurélio Zanchin, Nilson Ivo Tonin Santos, Fred Luciano Neves |
author_sort | Celedon, Paola Alejandra Fiorani |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8391164 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-83911642021-08-28 Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis Celedon, Paola Alejandra Fiorani Leony, Leonardo Maia Oliveira, Ueriton Dias Freitas, Natália Erdens Maron Silva, Ângelo Antônio Oliveira Daltro, Ramona Tavares Santos, Emily Ferreira Krieger, Marco Aurélio Zanchin, Nilson Ivo Tonin Santos, Fred Luciano Neves Biosensors (Basel) Article 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. MDPI 2021-08-22 /pmc/articles/PMC8391164/ /pubmed/34436091 http://dx.doi.org/10.3390/bios11080289 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Celedon, Paola Alejandra Fiorani Leony, Leonardo Maia Oliveira, Ueriton Dias Freitas, Natália Erdens Maron Silva, Ângelo Antônio Oliveira Daltro, Ramona Tavares Santos, Emily Ferreira Krieger, Marco Aurélio Zanchin, Nilson Ivo Tonin Santos, Fred Luciano Neves Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis |
title | Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis |
title_full | Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis |
title_fullStr | Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis |
title_full_unstemmed | Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis |
title_short | Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis |
title_sort | stability assessment of four chimeric proteins for human chagas disease immunodiagnosis |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8391164/ https://www.ncbi.nlm.nih.gov/pubmed/34436091 http://dx.doi.org/10.3390/bios11080289 |
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