Development of a Novel Glycoengineering Platform for the Rapid Production of Conjugate Vaccines

Overview

Journal Microb Cell Fact

Publisher Biomed Central

Specialties Biotechnology
Microbiology

Date 2023 Aug 18

PMID 37596672

Authors

Sherif Abouelhadid

Elizabeth R Atkins

Emily J Kay

Ian J Passmore

Simon J North

Burhan Lehri

Paul Hitchen

Eirik Bakke

Mohammed Rahman

Janine T Bosse

Yanwen Li

Vanessa S Terra

Paul R Langford

Anne Dell

Brendan W Wren

Jon Cuccui

Affiliations

Soon will be listed here.

Abstract

Conjugate vaccines produced either by chemical or biologically conjugation have been demonstrated to be safe and efficacious in protection against several deadly bacterial diseases. However, conjugate vaccine assembly and production have several shortcomings which hinders their wider availability. Here, we developed a tool, Mobile-element Assisted Glycoconjugation by Insertion on Chromosome, MAGIC, a novel biotechnological platform that overcomes the limitations of the current conjugate vaccine design method(s). As a model, we focused our design on a leading bioconjugation method using N-oligosaccharyltransferase (OTase), PglB. The installation of MAGIC led to at least twofold increase in glycoconjugate yield via MAGIC when compared to conventional N-OTase based bioconjugation method(s). Then, we improved MAGIC to (a) allow rapid installation of glycoengineering component(s), (b) omit the usage of antibiotics, (c) reduce the dependence on protein induction agents. Furthermore, we show the modularity of the MAGIC platform in performing glycoengineering in bacterial species that are less genetically tractable than the commonly used Escherichia coli. The MAGIC system promises a rapid, robust and versatile method to develop vaccines against serious bacterial pathogens. We anticipate the utility of the MAGIC platform could enhance vaccines production due to its compatibility with virtually any bioconjugation method, thus expanding vaccine biopreparedness toolbox.

Citing Articles

Recombinant production platform for Group A Streptococcus glycoconjugate vaccines.

Ajay Castro S, Passmore I, Ndeh D, Shaw H, Ruda A, Burns K NPJ Vaccines. 2025; 10(1):16.

PMID: 39843476 PMC: 11754613. DOI: 10.1038/s41541-025-01068-2.

A combinatorial DNA assembly approach to biosynthesis of N-linked glycans in E. coli.

Passmore I, Faulds-Pain A, Abouelhadid S, Harrison M, Hall C, Hitchen P Glycobiology. 2023; 33(2):138-149.

PMID: 36637423 PMC: 9990991. DOI: 10.1093/glycob/cwac082.

References

Rappuoli R . Glycoconjugate vaccines: Principles and mechanisms. Sci Transl Med. 2018; 10(456). DOI: 10.1126/scitranslmed.aat4615. View

Dow J, Mauri M, Scott T, Wren B . Improving protein glycan coupling technology (PGCT) for glycoconjugate vaccine production. Expert Rev Vaccines. 2020; 19(6):507-527. DOI: 10.1080/14760584.2020.1775077. View

Avci F, Li X, Tsuji M, Kasper D . A mechanism for glycoconjugate vaccine activation of the adaptive immune system and its implications for vaccine design. Nat Med. 2011; 17(12):1602-9. PMC: 3482454. DOI: 10.1038/nm.2535. View

Kay E, Cuccui J, Wren B . Recent advances in the production of recombinant glycoconjugate vaccines. NPJ Vaccines. 2019; 4:16. PMC: 6494827. DOI: 10.1038/s41541-019-0110-z. View

Poolman J, Frasch C, Nurkka A, Kayhty H, Biemans R, Schuerman L . Impact of the conjugation method on the immunogenicity of Streptococcus pneumoniae serotype 19F polysaccharide in conjugate vaccines. Clin Vaccine Immunol. 2010; 18(2):327-36. PMC: 3067356. DOI: 10.1128/CVI.00402-10. View

Ihssen J, Kowarik M, Dilettoso S, Tanner C, Wacker M, Thony-Meyer L . Production of glycoprotein vaccines in Escherichia coli. Microb Cell Fact. 2010; 9:61. PMC: 2927510. DOI: 10.1186/1475-2859-9-61. View

Cuccui J, Thomas R, Moule M, DElia R, Laws T, Mills D . Exploitation of bacterial N-linked glycosylation to develop a novel recombinant glycoconjugate vaccine against Francisella tularensis. Open Biol. 2013; 3(5):130002. PMC: 3866875. DOI: 10.1098/rsob.130002. View

Herbert J, Kay E, Faustini S, Richter A, Abouelhadid S, Cuccui J . Production and efficacy of a low-cost recombinant pneumococcal protein polysaccharide conjugate vaccine. Vaccine. 2018; 36(26):3809-3819. PMC: 5999350. DOI: 10.1016/j.vaccine.2018.05.036. View

Strutton B, Jaffe S, Pandhal J, Wright P . Producing a glycosylating Escherichia coli cell factory: The placement of the bacterial oligosaccharyl transferase pglB onto the genome. Biochem Biophys Res Commun. 2017; 495(1):686-692. DOI: 10.1016/j.bbrc.2017.11.023. View

10.

Dykxhoorn D, St Pierre R, Linn T . A set of compatible tac promoter expression vectors. Gene. 1996; 177(1-2):133-6. DOI: 10.1016/0378-1119(96)00289-2. View

11.

Oyston P, Sjostedt A, Titball R . Tularaemia: bioterrorism defence renews interest in Francisella tularensis. Nat Rev Microbiol. 2004; 2(12):967-78. DOI: 10.1038/nrmicro1045. View

12.

Ramachandran S, Deshpande O, Roseman C, Rosenberg N, Feldman M, Cavalli-Sforza L . Support from the relationship of genetic and geographic distance in human populations for a serial founder effect originating in Africa. Proc Natl Acad Sci U S A. 2005; 102(44):15942-7. PMC: 1276087. DOI: 10.1073/pnas.0507611102. View

13.

Weiser J, Ferreira D, Paton J . Streptococcus pneumoniae: transmission, colonization and invasion. Nat Rev Microbiol. 2018; 16(6):355-367. PMC: 5949087. DOI: 10.1038/s41579-018-0001-8. View

14.

Kay E, Yates L, Terra V, Cuccui J, Wren B . Recombinant expression of Streptococcus pneumoniae capsular polysaccharides in Escherichia coli. Open Biol. 2016; 6(4):150243. PMC: 4838161. DOI: 10.1098/rsob.150243. View

15.

Kelly J, Rubin A, Davis J, Ajo-Franklin C, Cumbers J, Czar M . Measuring the activity of BioBrick promoters using an in vivo reference standard. J Biol Eng. 2009; 3:4. PMC: 2683166. DOI: 10.1186/1754-1611-3-4. View

16.

Poli J, Guinoiseau E, de Rocca Serra D, Sutour S, Paoli M, Tomi F . Anti-Quorum Sensing Activity of 12 Essential Oils on and Specific Action of -Menthenolide from Corsican ssp. . Molecules. 2018; 23(9). PMC: 6225197. DOI: 10.3390/molecules23092125. View

17.

Rangel J, Sparling P, Crowe C, Griffin P, Swerdlow D . Epidemiology of Escherichia coli O157:H7 outbreaks, United States, 1982-2002. Emerg Infect Dis. 2005; 11(4):603-9. PMC: 3320345. DOI: 10.3201/eid1104.040739. View

18.

Vinogradov E, Conlan J, Perry M . Serological cross-reaction between the lipopolysaccharide O-polysaccharaide antigens of Escherichia coli O157:H7 and strains of Citrobcter freundii and Citrobacter sedlakii. FEMS Microbiol Lett. 2000; 190(1):157-61. DOI: 10.1111/j.1574-6968.2000.tb09279.x. View

19.

Bandyopadhyay A, Singh H, Fournier-Caruana J, Modlin J, Wenger J, Partridge J . Facility-Associated Release of Polioviruses into Communities-Risks for the Posteradication Era. Emerg Infect Dis. 2019; 25(7):1363-1369. PMC: 6590745. DOI: 10.3201/eid2507.181703. View

20.

Stefanetti G, Okan N, Fink A, Gardner E, Kasper D . Glycoconjugate vaccine using a genetically modified O antigen induces protective antibodies to . Proc Natl Acad Sci U S A. 2019; 116(14):7062-7070. PMC: 6452683. DOI: 10.1073/pnas.1900144116. View