Chapter 11
Biosynthesis of Glycoconjugate Virus-like Particles (VLPs)
Kathryn K. Oi, Tom A. Kloter, and Timothy G. Keys
Abstract
The outermost surface of bacterial pathogens consists primarily of complex carbohydrate structures—
polysaccharides, glycolipids, and glycoproteins. To raise a long-lasting and effective immune response
against carbohydrate antigens, they generally require covalent attachment to an immunogenic carrier
protein—a so-called glycoconjugate vaccine. One hurdle to the development of glycoconjugate vaccines
is that carbohydrate antigens remain inaccessible to recombinant production. Thus, the carbohydrate
antigen is typically purified from the pathogen and then chemically conjugated to an immunogenic protein.
Recent developments in the field of bacterial glycoengineering have opened the opportunity for total
recombinant production of glycoconjugate vaccines. In this method, we describe the production of
proteinaceous, virus-like particles (VLPs) bearing the conserved N-glycan of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumoniae.
Key words Actinobacillus pleuropneumoniae, N-glycosylation, Virus-like particle, Glycoconjugate,
Vaccine
1 Introduction
The rise of antibiotic-resistant bacterial pathogens threatens the
global health system. The root of this problem lies in our reliance
on antibiotics to control and prevent infection in clinical and agricultural settings. Vaccines play an important role in reducing the
demand for antibiotics; however, the design, production, and distribution of effective and economically viable vaccines still presents
significant technical challenges. Simple methods for producing
effective antibacterial vaccines at large-scale and low-cost are
urgently needed.
Actinobacillus pleuropneumoniae is the causative bacterial agent
of porcine pleuropneumonia—a disease underlying widespread
antibiotic use and economic losses in the pork industry. A range
of attenuated, inactivated, and subunit vaccines has been developed
against A. pleuropneumoniae [1]. However, due to their variable
surface structures (including lipopolysaccharide, capsular polysaccharides, and outer membrane proteins), none of the available
Blaine A. Pfeifer and Andrew Hill (eds.), Vaccine Delivery Technology: Methods and Protocols, Methods in Molecular Biology,
vol. 2183, https://doi.org/10.1007/978-1-0716-0795-4_11, © Springer Science+Business Media, LLC, part of Springer Nature 2021
205
Biosynthesis of Glycoconjugate Virus-like Particles (VLPs)
Kathryn K. Oi, Tom A. Kloter, and Timothy G. Keys
Abstract
The outermost surface of bacterial pathogens consists primarily of complex carbohydrate structures—
polysaccharides, glycolipids, and glycoproteins. To raise a long-lasting and effective immune response
against carbohydrate antigens, they generally require covalent attachment to an immunogenic carrier
protein—a so-called glycoconjugate vaccine. One hurdle to the development of glycoconjugate vaccines
is that carbohydrate antigens remain inaccessible to recombinant production. Thus, the carbohydrate
antigen is typically purified from the pathogen and then chemically conjugated to an immunogenic protein.
Recent developments in the field of bacterial glycoengineering have opened the opportunity for total
recombinant production of glycoconjugate vaccines. In this method, we describe the production of
proteinaceous, virus-like particles (VLPs) bearing the conserved N-glycan of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumoniae.
Key words Actinobacillus pleuropneumoniae, N-glycosylation, Virus-like particle, Glycoconjugate,
Vaccine
1 Introduction
The rise of antibiotic-resistant bacterial pathogens threatens the
global health system. The root of this problem lies in our reliance
on antibiotics to control and prevent infection in clinical and agricultural settings. Vaccines play an important role in reducing the
demand for antibiotics; however, the design, production, and distribution of effective and economically viable vaccines still presents
significant technical challenges. Simple methods for producing
effective antibacterial vaccines at large-scale and low-cost are
urgently needed.
Actinobacillus pleuropneumoniae is the causative bacterial agent
of porcine pleuropneumonia—a disease underlying widespread
antibiotic use and economic losses in the pork industry. A range
of attenuated, inactivated, and subunit vaccines has been developed
against A. pleuropneumoniae [1]. However, due to their variable
surface structures (including lipopolysaccharide, capsular polysaccharides, and outer membrane proteins), none of the available
Blaine A. Pfeifer and Andrew Hill (eds.), Vaccine Delivery Technology: Methods and Protocols, Methods in Molecular Biology,
vol. 2183, https://doi.org/10.1007/978-1-0716-0795-4_11, © Springer Science+Business Media, LLC, part of Springer Nature 2021
205