Single-Domain Antibody-Gold Nanoparticle Bioconjugates as Immunosensors for the Detection of Hantaviruses
- 26.05.2024
- Original Research Article
Erschienen in:
- Verfasst von
- Erika A. Bastos-Soares
- Michelle Suelen da Silva Morais
- Maribel Funes-Huacca
- Rosa Maria O. Sousa
- Nairo Brilhante-Da-Silva
- Sibele Andrade Roberto
- Nidiane Dantas R. Prado
- Claudia N. Duarte dos Santos
- Anna C. M. Marinho
- Andreimar M. Soares
- Rodrigo G. Stabeli
- Soraya dos Santos Pereira
- Carla Freire C. Fernandes
- Erschienen in
- Molecular Diagnosis & Therapy | Ausgabe 4/2024
Abstract
Introduction
Hantavirus, a zoonotic pathogen, causes severe syndromes like hemorrhagic fever with renal syndrome (HFRS), sometimes fatal in humans. Considering the importance of detecting the hantavirus antigen, the construction of an immunosensor is essential. The structural and functional characteristics of camelid nanobodies (VHHs) encourage their application in the areas of nanobiotechnology, therapeutics, diagnostics, and basic research. Therefore, this study aimed to standardize stable bioconjugates using gold nanoparticles (AuNPs) and VHHs, in order to develop immunobiosensors for the diagnosis of hantavirus infection.
Methods
Immobilized metal affinity chromatography (IMAC) was performed to obtain purified recombinant anti-hantavirus nucleocapsid nanobodies (anti-prNΔ85 VHH), while AuNPs were synthesized for bioconjugation. UV-visible spectrophotometry and transmission electron microscopy (TEM) analysis were employed to characterize AuNPs.
Results
The bioconjugation stability parameters (VHH-AuNPs), analyzed by spectrophotometry, showed that the ideal pH value and VHH concentration were obtained at 7.4 and 50 μg/mL, respectively, after addition of 1 M NaCl, which induces AuNP aggregation. TEM performed before and after bioconjugation showed uniform, homogeneous, well-dispersed, and spherical AuNPs with an average diameter of ~ 14 ± 0.57 nm. Furthermore, high-resolution images revealed a thin white halo on the surface of the AuNPs, indicating the coating of the AuNPs with protein. A biosensor simulation test (dot blot-like [DB-like]) was performed in stationary phase to verify the binding and detection limits of the recombinant nucleocapsid protein from the Araucária hantavirus strain (prN∆85).
Discussion
Using AuNPs/VHH bioconjugates, a specific interaction was detected between 5 and 10 min of reaction in a dose-dependent manner. It was observed that this test was sensitive enough to detect prNΔ85 at concentrations up to 25 ng/μL. Considering that nanostructured biological systems such as antibodies conjugated with AuNPs are useful tools for the development of chemical and biological sensors, the stability of the bioconjugate indicates proficiency in detecting antigens. The experimental results obtained will be used in a future immunospot assay or lateral flow immunochromatography analysis for hantavirus detection.
Graphical Abstract
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- Titel
- Single-Domain Antibody-Gold Nanoparticle Bioconjugates as Immunosensors for the Detection of Hantaviruses
- Verfasst von
-
Erika A. Bastos-Soares
Michelle Suelen da Silva Morais
Maribel Funes-Huacca
Rosa Maria O. Sousa
Nairo Brilhante-Da-Silva
Sibele Andrade Roberto
Nidiane Dantas R. Prado
Claudia N. Duarte dos Santos
Anna C. M. Marinho
Andreimar M. Soares
Rodrigo G. Stabeli
Soraya dos Santos Pereira
Carla Freire C. Fernandes
- Publikationsdatum
- 26.05.2024
- Verlag
- Springer International Publishing
- Erschienen in
-
Molecular Diagnosis & Therapy / Ausgabe 4/2024
Print ISSN: 1177-1062
Elektronische ISSN: 1179-2000 - DOI
- https://doi.org/10.1007/s40291-024-00713-1
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