title: Scalable chromatography-based purification of virus-like particle carrier for epitope based influenza A vaccine produced in Escherichia coli.
abstract: Virus-like particles (VLPs) are promising molecular structures for the design and construction of novel vaccines, diagnostic tools, and gene therapy vectors. Size, oligomer assembly and repetitiveness of epitopes are optimal features to induce strong immune responses. Several VLP-based vaccines are currently licensed and commercialized, and many vaccine candidates are now under preclinical and clinical studies. In recent years, the development of genetically engineered recombinant VLPs has accelerated the need for new, improved downstream processes. In particular, a rapid low cost purification process has been identified as a remaining key challenge in manufacturing process development. In the present study we set up a size-exclusion chromatography-based, scalable purification protocol for the purification of a VLP-based influenza A vaccine produced in Escherichia coli. Recombinant VLPs derived from the RNA bacteriophage MS2 displaying an epitope from the ectodomain of Matrix 2 protein from influenza A virus were produced and purified. The 3 steps purification protocol uses a recently developed multimodal size-exclusion chromatography medium (Capto™ Core 700) in combination with detergent extraction and size-exclusion polishing to reach a 89% VLP purity with a 19% yield. The combination of this downstream strategy following production in E. coli would be suited for production of VLP-based veterinary vaccines targeting livestock and companion animals where large amounts of doses must be produced at an affordable price.
authors.affiliation: BIOASTER, Protein and Expression System Engineering unit, 40 avenue Tony Garnier, 69007 Lyon, France.
authors.forename: Priscillia
authors.initials: P
authors.lastname: Lagoutte
authors.affiliation: BIOASTER, Protein and Expression System Engineering unit, 40 avenue Tony Garnier, 69007 Lyon, France.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Mignon
authors.affiliation: BIOASTER, Protein and Expression System Engineering unit, 40 avenue Tony Garnier, 69007 Lyon, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Donnat
authors.affiliation: BIOASTER, Protein and Expression System Engineering unit, 40 avenue Tony Garnier, 69007 Lyon, France.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Stadthagen
authors.affiliation: EM Service, CODA-CERVA, Groeselenbergstraat 99, B-1180 Brussels, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Mast
authors.affiliation: BIOASTER, Protein and Expression System Engineering unit, 40 avenue Tony Garnier, 69007 Lyon, France.
authors.forename: Régis
authors.initials: R
authors.lastname: Sodoyer
authors.affiliation: BIOASTER, Protein and Expression System Engineering unit, 40 avenue Tony Garnier, 69007 Lyon, France. Electronic address: adrien.lugari@bioaster.org.
authors.forename: Adrien
authors.initials: A
authors.lastname: Lugari
authors.affiliation: BIOASTER, Protein and Expression System Engineering unit, 40 avenue Tony Garnier, 69007 Lyon, France. Electronic address: bettina.werle@bioaster.org.
authors.forename: Bettina
authors.initials: B
authors.lastname: Werle
mesh_terms: D000818:Animals; D002850:Chromatography, Gel / Q000379:methods*; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009980:Influenza A virus / Q000235:genetics / Q000276:immunology*; D007252:Influenza Vaccines / Q000235:genetics / Q000302:isolation & purification*; D017909:Levivirus / Q000235:genetics; D009976:Orthomyxoviridae Infections / Q000517:prevention & control / Q000662:veterinary; D014614:Vaccines, Synthetic / Q000235:genetics / Q000302:isolation & purification; D058425:Vaccines, Virus-Like Particle / Q000235:genetics / Q000302:isolation & purification*; D014730:Veterinary Medicine / Q000379:methods; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology; D000086082:Viroporin Proteins
chemical_list: D000939:Epitopes; D007252:Influenza Vaccines; D014614:Vaccines, Synthetic; D058425:Vaccines, Virus-Like Particle; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins
keywords: Downstream process; Liquid chromatography; Purification; VLPs; Virus-like particles