Efficient virus detection utilizing chitin-immobilized nanobodies synthesized in Ustilago maydis.
Chitin
Chitinase
Nanobody
SARS-CoV-2
Ustilago maydis
Journal
Journal of biotechnology
ISSN: 1873-4863
Titre abrégé: J Biotechnol
Pays: Netherlands
ID NLM: 8411927
Informations de publication
Date de publication:
20 Mar 2023
20 Mar 2023
Historique:
received:
12
12
2022
revised:
08
02
2023
accepted:
16
03
2023
medline:
4
4
2023
pubmed:
23
3
2023
entrez:
22
3
2023
Statut:
ppublish
Résumé
The COVID-19 pandemic has greatly impacted the global economy and health care systems, illustrating the urgent need for timely and inexpensive responses to pandemic threats in the form of vaccines and antigen tests. Currently, antigen testing is mostly conducted by qualitative flow chromatography or via quantitative ELISA-type assays. The latter mostly utilize materials like protein-adhesive polymers and gold or latex particles. Here we present an alternative ELISA approach using inexpensive, biogenic materials and permitting quick detection based on components produced in the microbial model Ustilago maydis. In this fungus, heterologous proteins like biopharmaceuticals can be exported by fusion to unconventionally secreted chitinase Cts1. As a unique feature, the carrier chitinase binds to chitin allowing its additional use as a purification or immobilization tag. Recent work has demonstrated that nanobodies are suitable target proteins. These proteins represent a very versatile alternative antibody format and can quickly be adapted to detect novel antigens by camelidae immunization or synthetic libraries. In this study, we exemplarily produced different mono- and bivalent SARS-CoV-2 nanobodies directed against the spike protein receptor binding domain (RBD) as Cts1 fusions and screened their antigen binding affinity in vitro and in vivo. Functional nanobody-Cts1 fusions were immobilized on chitin forming an RBD tethering surface. This provides a solid base for future development of inexpensive antigen tests utilizing unconventionally secreted nanobodies as antigen trap and a matching ubiquitous and biogenic surface for immobilization.
Identifiants
pubmed: 36948402
pii: S0168-1656(23)00057-3
doi: 10.1016/j.jbiotec.2023.03.005
pmc: PMC10028217
pii:
doi:
Substances chimiques
Single-Domain Antibodies
0
Chitin
1398-61-4
Chitinases
EC 3.2.1.14
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
72-84Informations de copyright
Copyright © 2023 Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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