development of a novel glycoengineering platform for the rapid production of conjugate CORD-Papers-2022-06-02 (Version 1)

Title: Development of a novel glycoengineering platform for the rapid production of conjugate vaccines
Abstract: Antimicrobial resistance (AMR) is threatening the lives of millions worldwide. Antibiotics which once saved countless lives are now failing ushering in vaccines development as a current global imperative. Conjugate vaccines produced either by chemical synthesis or biologically in Escherichia coli cells have been demonstrated to be safe and efficacious in protection against several deadly bacterial diseases. However conjugate vaccines 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 method that overcomes the limitations of the current conjugate vaccine design method(s). We demonstrate at least 2-fold increase in glycoconjugate yield via MAGIC when compared to conventional bioconjugate method(s). Furthermore the modularity of the MAGIC platform also allowed us to perform glycoengineering in genetically intractable bacterial species other than E. coli. The MAGIC system promises a rapid robust and versatile method to develop vaccines against bacteria especially AMR pathogens and could be applied for biopreparedness.
Published: 2021-11-25
Journal: bioRxiv
DOI: 10.1101/2021.11.25.470047
DOI_URL: http://doi.org/10.1101/2021.11.25.470047
Author Name: Abouelhadid Sherif
Author link: https://covid19-data.nist.gov/pid/rest/local/author/abouelhadid_sherif
Author Name: Atkins Elizabeth
Author link: https://covid19-data.nist.gov/pid/rest/local/author/atkins_elizabeth
Author Name: Kay Emily
Author link: https://covid19-data.nist.gov/pid/rest/local/author/kay_emily
Author Name: Passmore Ian
Author link: https://covid19-data.nist.gov/pid/rest/local/author/passmore_ian
Author Name: North Simon J
Author link: https://covid19-data.nist.gov/pid/rest/local/author/north_simon_j
Author Name: Lehri Burhan
Author link: https://covid19-data.nist.gov/pid/rest/local/author/lehri_burhan
Author Name: Hitchen Paul
Author link: https://covid19-data.nist.gov/pid/rest/local/author/hitchen_paul
Author Name: Bakke Eirik
Author link: https://covid19-data.nist.gov/pid/rest/local/author/bakke_eirik
Author Name: Rahman Mohammed
Author link: https://covid19-data.nist.gov/pid/rest/local/author/rahman_mohammed
Author Name: Bosse Janine
Author link: https://covid19-data.nist.gov/pid/rest/local/author/bosse_janine
Author Name: Li Yanwen
Author link: https://covid19-data.nist.gov/pid/rest/local/author/li_yanwen
Author Name: Terra Vanessa S
Author link: https://covid19-data.nist.gov/pid/rest/local/author/terra_vanessa_s
Author Name: Langford Paul
Author link: https://covid19-data.nist.gov/pid/rest/local/author/langford_paul
Author Name: Dell Anne
Author link: https://covid19-data.nist.gov/pid/rest/local/author/dell_anne
Author Name: Wren Brendan W
Author link: https://covid19-data.nist.gov/pid/rest/local/author/wren_brendan_w
Author Name: Cuccui Jon
Author link: https://covid19-data.nist.gov/pid/rest/local/author/cuccui_jon
sha: c24f5b5f444b822a8a66772c0325883b989278cf
license: biorxiv
license_url: https://www.biorxiv.org/about-biorxiv
source_x: BioRxiv
source_x_url: https://www.biorxiv.org/
url: https://doi.org/10.1101/2021.11.25.470047
has_full_text: TRUE
Keywords Extracted from Text Content: glycoconjugate Escherichia coli cells AMR E. coli recombinase human pELLA2 OST chromosomal S. pneumoniae serotype 4 OST PglB E. coli strains cellular people pEXT20 pglBx15 Mini-Tn5Km2 Vaccines T-cell glycoconjugates S30 buffer vaccine micro-factory children pglB PCV13 E. coli S. pneumoniae glycoconjugate E. coli cell mouse SP4 component(s IPTG trypsin or chymotrypsin glycoconjugate vaccines dithiothreitol pELLA3 S. pneumoniae tac line SfiI Sp4 carbohydrate moiety NotI E. coli W3110 CmeA-Sp4 glycoconjugates (29) MI4A Trypsin C. jejuni NCTC11168 PglB plugand-play LLO C. jejuni pglB cell n-dodecylβ-d-maltopyranoside Cell DDM pUTminiTn5 glycans plug- glycoconjugate glycoconjugate vaccine E. coli O157 outbred mice Biotechnology U169 deoRrecA1 endA1 hsdR17 BB/M01925X/1 E. coli Lynham F-φ80lacZΔM15 Δ(lacZYA-argF BB/H017437/1 UK λ-thi-1 gyrA96 relA1
Extracted Text Content in Record: First 5000 Characters:Antimicrobial resistance (AMR) is threatening the lives of millions worldwide. Antibiotics which once saved countless lives, are now failing, ushering in vaccines development as a current global imperative. Conjugate vaccines produced either by chemical synthesis or biologically in Escherichia coli cells, have been demonstrated to be safe and efficacious in protection against several deadly bacterial diseases. However, conjugate vaccines 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 method that overcomes the limitations of the current conjugate vaccine design method(s). We demonstrate at least 2-fold increase in glycoconjugate yield via MAGIC when compared to conventional bioconjugate method(s). Furthermore, the modularity of the MAGIC platform also allowed us to perform glycoengineering in genetically intractable bacterial species other than E. coli. The MAGIC system promises a rapid, robust and versatile method to develop vaccines against bacteria, especially AMR pathogens, and could be applied for biopreparedness. The alarming rise in antimicrobial resistance necessitates global efforts to prevent a future health crisis. For more than half a century, antibiotics were considered the first line of defence against bacterial pathogens (1) . However, the spread of antibiotic resistance amongst pathogenic bacteria entails considerable efforts to look for antibiotic alternatives. Vaccines have been successful in curbing infectious diseases for decades, not only among adults but also among children and the elderly, thus saving millions of lives worldwide (2) . According to the Market Information for Access to Vaccines (MI4A), World Health Organization, the vaccines market is estimated to be worth approximately $33 billion in 2019 (3). Current biotechnological platforms however, might not be able to fulfil the vaccines supply demand. To satisfy the market's demand for conjugate vaccines, to protect humanity from a foreseeable pandemic, and to be able to tailor novel efficacious vaccines at lower cost, significant biotechnological innovation is needed. Glycoconjugate vaccines are considered to be one of the safest and most effective tools to combat serious infectious diseases including bacterial meningitis and pneumonia (4) . Conjugation is achieved by linking glycans (carbohydrate moiety), either chemically or enzymatically, to proteins via covalent bonds. This leads to a T-cell dependent immune response, offering excellent protection in people of all ages (5) . Traditionally chemical approaches to produce glycoconjugate vaccine involve the activation of functional groups on the glycan and protein that are linked chemically in a multi-step method that is expensive and laborious, requiring several rounds of purification after each step (6) . Additionally, chemical conjugation methods such as reductive amination can alter the polysaccharide epitope, affecting the immunogenicity of the glycoconjugate against the disease, besides its inherent batch-to-batch variation (7) . Biological conjugation (bioconjugation) offers an excellent alternative to chemical conjugation. It is based on using a bacterial cell, usually E. coli, as a chassis to express a pathway that encodes the desired bacterial polysaccharide, carrier protein, and an oligosaccharytransferase enzyme, OST, that catalyses the conjugation process (6) . The advent of the bacterial bioconjugation method allowed several protein glycan vaccine combinations to be successfully developed, emphasizing its immense potential to become the preferred method to develop glycoconjugate vaccines in the future (4, (8) (9) (10) (11) (12) . However, several challenges remain. Firstly, the process places significant metabolic stress on the E. coli cell, vaccine micro-factory, due to the expression of orthogonal pathways (13) . This process requires the prior genetic and structural information of the polysaccharide structure of choice. Secondly, the use of three independent replicons has limitations due to incompatibility of plasmid origins of replication and antibiotic selection markers which may lead to the plasmid loss that results in reduction in glycoconjugate yield (8, 10, 14) . Thirdly, reports have demonstrated that the expression of the OST PglB, that catalyses the linking of glycans to carrier proteins, has a detrimental effect on bacterial growth, thus decreasing cellular fitness to produce glycoconjugates (13, 15) . All this together results in a low biomass which often translates to a reduction in the vaccine yield. Consequently, this leads to an increase in the production cost of a glycoconjugate vaccine, making it unaffordable in low-income countries where they are most needed, putting millions of lives at risk as a result of vaccines inequity (6) . Previous attempts to engineer robust glycoengineering host stra
PDF JSON Files: document_parses/pdf_json/c24f5b5f444b822a8a66772c0325883b989278cf.json
G_ID: development_of_a_novel_glycoengineering_platform_for_the_rapid_production_of_conjugate