TY - JOUR T1 - Scalable chromatography-based purification of virus-like particle carrier for epitope based influenza A vaccine produced in Escherichia coli. JF - J Virol Methods Y1 - 2016 A1 - Lagoutte, Priscillia A1 - Mignon, Charlotte A1 - Donnat, Stéphanie A1 - Stadthagen, Gustavo A1 - Jan Mast A1 - Sodoyer, Régis A1 - Lugari, Adrien A1 - Werle, Bettina KW - Animals KW - Chromatography, Gel KW - Epitopes KW - Escherichia coli KW - Influenza A virus KW - Influenza Vaccines KW - Levivirus KW - Orthomyxoviridae Infections KW - Vaccines, Synthetic KW - Vaccines, Virus-Like Particle KW - Veterinary Medicine KW - Viral Matrix Proteins AB -

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.

VL - 232 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26947397?dopt=Abstract M3 - 10.1016/j.jviromet.2016.02.011 ER -