Development of an automated platform for the optimal production of glycoconjugate vaccines expressed in Escherichia coli

• Published in: Microbial cell factories Vol. 20; no. 1; pp. 104 - 15
• Main Authors: Samaras, Jasmin J., Mauri, Marta, Kay, Emily J., Wren, Brendan W., Micheletti, Martina
• Format: Journal Article
• Language: English
• Published: London BioMed Central 24.05.2021; BioMed Central Ltd; BMC
• Subjects: ADP Ribose Transferases - metabolism; Antibiotics; Antigens; Applications software; Applied Microbiology; Automation; Automation - methods; Bacterial Toxins - metabolism; Bacterial Vaccines - biosynthesis; Bioconjugation; Biotechnology; Biotechnology - methods; Cell culture; Chemistry; Chemistry and Materials Science; Clinical trials; Collaboration; Computer applications; Conjugate vaccines; Conjugation; E coli; Enzymes; Enzymology; Epidemics; Escherichia coli; Escherichia coli - metabolism; Exotoxins - metabolism; Genetic aspects; Genetic Engineering; Glycan; Glycoconjugate vaccine; Glycosylation; High-Throughput Screening Assays - methods; Humans; Infectious diseases; Methods; Microbial genetic engineering; Microbial Genetics and Genomics; Microbial glycobiotechnology; Microbiological research; Microbiology; Peptides; PGCT; Physiological aspects; Plasmids; Pneumococcal vaccine; Pneumococcal Vaccines - biosynthesis; Pneumonia; Polysaccharides; Polysaccharides, Bacterial - metabolism; Production processes; Protein engineering; Proteins; Pseudomonas aeruginosa Exotoxin A; R&D; Research & development; Robustness (mathematics); Screening studies; Severe acute respiratory syndrome coronavirus 2; Streptococcus infections; Technology, Pharmaceutical - methods; Vaccines; Vaccines, Conjugate - biosynthesis; Virulence Factors - metabolism; Denmark; Europe; Automation; Glycoprotein; Glycoengineering; Glycoconjugate vaccine; Bioconjugation; High throughput process development; Screening studies; Pneumococcal vaccine; PGCT
• ISSN: 1475-2859; 1475-2859
• DOI: 10.1186/s12934-021-01588-1

Protein Glycan Coupling Technology (PGCT) uses purposely modified bacterial cells to produce recombinant glycoconjugate vaccines. This vaccine platform holds great potential in this context, namely due to its modular nature, the simplified production process in comparison to traditional chemical conjugation methods, and its amenability to scaled-up operations. As a result, a considerable reduction in production time and cost is expected, making PGCT-made vaccines a suitable vaccine technology for low-middle income countries, where vaccine coverage remains predominantly low and inconsistent. This work aims to develop an integrated whole-process automated platform for the screening of PGCT-made glycoconjugate vaccine candidates. The successful translation of a bench scale process for glycoconjugate production to a microscale automated setting was achieved. This was integrated with a numerical computational software that allowed hands-free operation and a platform adaptable to biological variation over the course of a production process. Platform robustness was proven with both technical and biological replicates and subsequently the platform was used to screen for the most favourable conditions for production of a pneumococcal serotype 4 vaccine candidate. This work establishes an effective automated platform that enabled the identification of the most suitable E. coli strain and genetic constructs to be used in ongoing early phase research and be further brought into preclinical trials.