Development of an O-polysaccharide based recombinant glycoconjugate vaccine in engineered E. coli against ExPEC O1

Extraintestinal pathogenic Escherichia coli O1 is a frequently identified serotype that causes serious infections and is often refractory to antimicrobial therapy. Glycoconjugate vaccine represents a promising measure to reduce ExPEC infections. Herein, we designed an O1-specific glyco-optimized chassis strain for manufacture of Opolysaccharide (OPS) antigen and OPS-based bioconjugate. Specifically, OPS and OPS-based glycoprotein were synthesized in glyco-optimized chassis strain, when compared to the unmeasurable level of the parent strain. The optimal expression of oligosaccharyltransferase and carrier protein further improved the titer. MS analysis elucidated the correct structure of resulting bioconjugate at routine and unreported glycosylation sequons of carrier protein, with a higher glycosylation efficiency. Finally, purified bioconjugate stimulated mouse to generate specific IgG antibodies and protected them against virulent ExPEC O1 challenge. The plug-and-play glyco-optimized platform is suitable for bioconjugate synthesis, thus providing a potential platform for future medical applications.

Automated summary

ExPEC O1 is a common pathogen causing serious infections and often resistant to antibiotics. A glycoconjugate vaccine was designed to target this strain, with promising results in producing OPS antigen and bioconjugates in a glyco-optimized chassis strain, leading to successful generation of specific IgG antibodies and protection in mice against ExPEC O1 challenge. This plug-and-play glyco-optimized platform shows potential for future medical applications.