Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Design
2.2. IBMP Protein Expression and Purification
2.3. Gel Electrophoresis
2.4. Circular Dichroism (CD) Measurements
2.5. Dynamic Light Scattering (DLS)
2.6. Clinical Specimens and Indirect ELISA
2.7. Data Analysis
3. Results
3.1. Reversibility of Heat-Induced Denaturation
3.2. Storage Stability at 25 °C
3.3. Long-Term Stability Analysis at 4 °C
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Chimeric Antigen | Sequence Name | Amino Acid Range | Gene Bank Sequence ID |
---|---|---|---|
IBMP-8.1 | Trans-sialidase | 747–774 | XP_820062.1 |
60S ribosomal protein L19 | 218–238 | XP_820995.1 | |
Trans-sialidase | 1435–1449 | XP_813586.1 | |
Surface antigen 2 (CA-2) | 276–297 | XP_813516.1 | |
IBMP-8.2 | Antigen, partial | 13–73 | ACM47959.1 |
Surface antigen 2 (CA-2) | 166–220 | XP_818927.1 | |
Calpain cysteine peptidase | 31–97 | XP_804989.1 | |
IBMP-8.3 | Trans-sialidase | 710–754 | XP_813237.1 |
Flagellar repetitive antigen protein | 15–56 | AAA30177.1 | |
60S ribosomal protein L19 | 236–284 | XP_808122.1 | |
Surface antigen 2 (CA-2) | 279–315 | XP_813516.1 | |
IBMP-8.4 | Shed-acute-phase-antigen | 681–704 | CAA40511.1 |
Kinetoplastid membrane protein KMP-11 | 76–92 | XP_810488.1 | |
Trans-sialidase | 1436–1449 | XP_813586.1 | |
Flagellar repetitive antigen protein | 20–47 | AAA30177.1 | |
Trans-sialidase | 740–759 | XP_820062.1 | |
Surface antigen 2 (CA-2) | 276–298 | XP_813516.1 | |
Flagellar repetitive antigen protein | 1–68 | AAA30197.1 | |
60S ribosomal protein L19 | 218–238 | XP_820995.1 | |
Microtubule-associated protein | 421–458 | XP_809567.1 |
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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
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 StyleCeledon, Paola Alejandra Fiorani, Leonardo Maia Leony, Ueriton Dias Oliveira, Natália Erdens Maron Freitas, Ângelo Antônio Oliveira Silva, Ramona Tavares Daltro, Emily Ferreira Santos, Marco Aurélio Krieger, Nilson Ivo Tonin Zanchin, and Fred Luciano Neves Santos. 2021. "Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis" Biosensors 11, no. 8: 289. https://doi.org/10.3390/bios11080289