title: Characterization of epitope-tagged foot-and-mouth disease virus.
abstract: Foot-and-mouth disease (FMD) is a highly contagious and economically devastating disease of cloven-hoofed animals with an almost-worldwide distribution. Conventional FMD vaccines consisting of chemically inactivated viruses have aided in the eradication of FMD from Europe and remain the main tool for control in endemic countries. Although significant steps have been made to improve the quality of vaccines, such as improved methods of antigen concentration and purification, manufacturing processes are technically demanding and expensive. Consequently, there is large variation in the quality of vaccines distributed in FMD-endemic countries compared with those manufactured for emergency use in FMD-free countries. Here, we have used reverse genetics to introduce haemagglutinin (HA) and FLAG tags into the foot-and-mouth disease virus (FMDV) capsid. HA- and FLAG-tagged FMDVs were infectious, with a plaque morphology similar to the non-tagged parental infectious copy virus and the field virus. The tagged viruses utilized integrin-mediated cell entry and retained the tag epitopes over serial passages. In addition, infectious HA- and FLAG-tagged FMDVs were readily purified from small-scale cultures using commercial antibodies. Tagged FMDV offers a feasible alternative to the current methods of vaccine concentration and purification, a potential to develop FMD vaccine conjugates and a unique tool for FMDV research.
authors: Pirbright Laboratory, Institute for Animal Health, Woking, Surrey, GU24 0NF, UK.
authors: Julian
authors: J
authors: Seago
authors: Pirbright Laboratory, Institute for Animal Health, Woking, Surrey, GU24 0NF, UK.
authors: Terry
authors: T
authors: Jackson
authors: Pirbright Laboratory, Institute for Animal Health, Woking, Surrey, GU24 0NF, UK.
authors: Claudia
authors: C
authors: Doel
authors: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors: Elizabeth
authors: E
authors: Fry
authors: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors: David
authors: D
authors: Stuart
authors: Central Veterinary Institute Wageningen UR, Division Virology, PO Box 65, 8200 AB Lelystad, The Netherlands.
authors: Michiel M
authors: MM
authors: Harmsen
authors: Pirbright Laboratory, Institute for Animal Health, Woking, Surrey, GU24 0NF, UK.
authors: Bryan
authors: B
authors: Charleston
authors: Pirbright Laboratory, Institute for Animal Health, Woking, Surrey, GU24 0NF, UK.
authors: Nicholas
authors: N
authors: Juleff
mesh_terms: D000818:Animals; D000914:Antibodies, Viral; D015153:Blotting, Western; D036022:Capsid Proteins / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005536:Foot-and-Mouth Disease / Q000517:prevention & control* / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000145:classification*; D008958:Models, Molecular; D011487:Protein Conformation; D012367:RNA, Viral; D013194:Staining and Labeling; D019444:Vaccines, DNA; D015164:Vaccines, Inactivated; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D000939:Epitopes; D012367:RNA, Viral; C442501:VP1 protein, Foot-and-mouth disease virus; D019444:Vaccines, DNA; D015164:Vaccines, Inactivated; D014765:Viral Vaccines