Semiautomated design and soluble expression of a chimeric antigen TbpAB01 from Glaesserella parasuis.
Biotechnol Bioeng
; 121(7): 2163-2174, 2024 Jul.
Article
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| MEDLINE
| ID: mdl-38595326
ABSTRACT
Pathogenic bacterial membrane proteins (MPs) are a class of vaccine and antibiotic development targets with widespread clinical application. However, the inherent hydrophobicity of MPs poses a challenge to fold correctly in living cells. Herein, we present a comprehensive method to improve the soluble form of MP antigen by rationally designing multi-epitope chimeric antigen (ChA) and screening two classes of protein-assisting folding element. The study uses a homologous protein antigen as a functional scaffold to generate a ChA possessing four epitopes from transferrin-binding protein A of Glaesserella parasuis. Our engineered strain, which co-expresses P17 tagged-ChA and endogenous chaperones groEL-ES, yields a 0.346 g/L highly soluble ChA with the property of HPS-positive serum reaction. Moreover, the protein titer of ChA reaches 4.27 g/L with >90% soluble proportion in 5-L bioreactor, which is the highest titer reported so far. The results highlight a timely approach to design and improve the soluble expression of MP antigen in industrially viable applications.
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Base de datos:
MEDLINE
Asunto principal:
Antígenos Bacterianos
Idioma:
En
Revista:
Biotechnol Bioeng
Año:
2024
Tipo del documento:
Article