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oai:figshare.com:article/33949615 2026-09-21T05:29:39Z category_696 portal_316 item_type_3 month_year_09_2026

Supplementary file 1_Development and evaluation of a dual-target glycoconjugate against Shigella sonnei.docx Catherine L. Hall (25080367) Tom L. Flood (25080370) Simon Clare (125268) Katherine Harcourt (488197) Emily J. Kay (10868576) Stephen Baker (30387) Brendan W. Wren (6750428) Genetic Immunology bioconjugate glycoconjugate O-antigen Shigella Shigella-specific protein shigellosis vaccine glycoengineering Introduction<p>Shigellosis morbidity and mortality, combined with the increase in multidrug-resistant infections, make Shigella vaccine development a global imperative. Glycoconjugate vaccines that couple immunogenic O-antigen to protein derived from Shigella may provide broader protection across Shigella species and serogroups. Such an approach also circumvents immunotolerance arising from repeated use of the same carrier. Here we use bioconjugation, exploiting an oligosaccharyltransferase (OST) enzyme to couple O-antigen and carrier protein in vivo, to generate a Shigella-specific dual-target glycoconjugate.</p>Methods<p>Glycoconjugates were synthesised in E. coli SDB1 cells producing S. sonnei O-antigen, the OST PglS, and one of two Shigella carrier proteins or the standard bioconjugate carrier, ExoA. Recombinant glycoconjugate was purified using anion exchange chromatography and then used to immunise mice. Antibody responses were measured and compared by ELISA. </p>Results<p>When co-produced in E. coli, PglS was able to transfer the cloned S. sonnei O-antigen onto three carrier proteins, modified to accept glycans from the PglS transferase enzyme—ExoA and two immunogenic Shigella-specific outer membrane proteins, EmrK and MdtA. Production of MdtA or ExoA glycoconjugates for immunisation studies utilised successive rounds of anion exchange chromatography, to remove unglycosylated material and obtain highly purified glycosylated proteins. Analysis of murine sera following immunisation revealed an IgG response was raised against both carrier protein and the S. sonnei O-antigen for each glycoconjugate. </p>Discussion<p>A novel, conserved Shigella protein can be utilised as an effective carrier for the S. sonnei O-antigen to generate a Shigella-specific glycoconjugate that elicits IgG responses to both carrier protein and S. sonnei O-antigen.</p> 2026-09-21T05:29:39Z Dataset Dataset 10.3389/fimmu.2026.1907725.s001 https://figshare.com/articles/dataset/Supplementary_file_1_Development_and_evaluation_of_a_dual-target_glycoconjugate_against_Shigella_sonnei_docx/33949615 CC BY 4.0