title: Non-chromatographic preparation of a bacterially produced single-shot modular virus-like particle capsomere vaccine for avian influenza.
abstract: Highly pathogenic avian influenza (HPAI) causes significant economic loss, reduced food security and poses an ongoing pandemic threat. Poultry vaccination significantly decreases these problems and recognizes that the health of humans, animals and ecosystems are connected. Low-cost manufacture of poultry vaccine matched quickly to the ever-changing circulating strain is needed for effective vaccination. Here, we re-engineered the process to manufacture bacterially synthesized modular capsomere comprising influenza M2e, previously shown to confer complete protection in challenged mice, for application in poultry. Modular capsomere was prepared using a simplified non-chromatographic salting-out precipitation method and its immunogenicity tested in vivo in poultry. Modular capsomere crudely purified by precipitation (pCapM2e) contained more contaminants than equivalent product purified by chromatography (cCapM2e). Unadjuvanted pCapM2e containing 80 EU of endotoxin per dose was inferior to highly purified and adjuvanted cCapM2e (2 EU per dose). However, addition of adjuvant to pCapM2e resulting in high immunogenicity after only a single dose of vaccination, yet without any local adverse reaction. This finding suggests a strong synergy between adjuvant, antigen and contaminants, and the possible existence of a "Goldilocks" level of contaminants, where high immunogenicity and low reactogenicity can be obtained in a single-shot vaccination. The simplified process offers potential cost and speed advantages to address the needs in influenza poultry vaccination in low-cost veterinary markets.
authors.affiliation: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, St Lucia, QLD 4072, Australia.
authors.forename: Nani
authors.initials: N
authors.lastname: Wibowo
authors.affiliation: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, St Lucia, QLD 4072, Australia.
authors.forename: Yang
authors.initials: Y
authors.lastname: Wu
authors.affiliation: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, St Lucia, QLD 4072, Australia.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Fan
authors.affiliation: The University of Queensland, School of Veterinary Science, Gatton, QLD 4343, Australia.
authors.forename: Joanne
authors.initials: J
authors.lastname: Meers
authors.affiliation: The University of Queensland, Protein Expression Facility, St Lucia, QLD 4072, Australia.
authors.forename: Linda H L
authors.initials: LH
authors.lastname: Lua
authors.affiliation: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, St Lucia, QLD 4072, Australia. Electronic address: a.middelberg@uq.edu.au.
authors.forename: Anton P J
authors.initials: AP
authors.lastname: Middelberg
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D011232:Chemical Precipitation; D002645:Chickens; D064420:Drug-Related Side Effects and Adverse Reactions / Q000453:epidemiology / Q000473:pathology; D007074:Immunoglobulin G / Q000097:blood; D005585:Influenza in Birds / Q000276:immunology / Q000517:prevention & control*; D051379:Mice; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D058425:Vaccines, Virus-Like Particle / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D014763:Viral Matrix Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D000086082:Viroporin Proteins
chemical_list: D000276:Adjuvants, Immunologic; D000914:Antibodies, Viral; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D058425:Vaccines, Virus-Like Particle; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins
keywords: Capsomere; Influenza; Polyomavirus; Poultry; Vaccine