title: Application of the thermofluor PaSTRy technique for improving foot-and-mouth disease virus vaccine formulation.
abstract: Foot-and-mouth disease (FMD) has a major economic impact throughout the world and is a considerable threat to food security. Current FMD virus (FMDV) vaccines are made from chemically inactivated virus and need to contain intact viral capsids to maximize efficacy. FMDV exists as seven serotypes, each made up by a number of constantly evolving subtypes. A lack of immunological cross-reactivity between serotypes and between some strains within a serotype greatly complicates efforts to control FMD by vaccination. Thus, vaccines for one serotype do not afford protection against the others, and multiple-serotype-specific vaccines are required for effective control. The FMDV serotypes exhibit variation in their thermostability, and the capsids of inactivated preparations of the O, C and SAT serotypes are particularly susceptible to dissociation at elevated temperature. Methods to quantify capsid stability are currently limited, lack sensitivity and cannot accurately reflect differences in thermostability. Thus, new, more sensitive approaches to quantify capsid stability would be of great value for the production of more stable vaccines and to assess the effect of production conditions on vaccine preparations. Here we have investigated the application of a novel methodology (termed PaSTRy) that utilizes an RNA-binding fluorescent dye and a quantitative (q)PCR machine to monitor viral genome release and hence dissociation of the FMDV capsid during a slow incremental increase in temperature. PaSTRy was used to characterize capsid stability of all FMDV serotypes. Furthermore, we have used this approach to identify stabilizing factors for the most labile FMDV serotypes.
authors.affiliation: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: Abhay
authors.initials: A
authors.lastname: Kotecha
authors.affiliation: The Pirbright Institute, Woking, Surrey, GU24 0NF, UK.
authors.forename: Fuquan
authors.initials: F
authors.lastname: Zhang
authors.affiliation: The Pirbright Institute, Woking, Surrey, GU24 0NF, UK.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Juleff
authors.affiliation: The Pirbright Institute, Woking, Surrey, GU24 0NF, UK.
authors.forename: Terry
authors.initials: T
authors.lastname: Jackson
authors.affiliation: The Pirbright Institute, Woking, Surrey, GU24 0NF, UK.
authors.forename: Eva
authors.initials: E
authors.lastname: Perez
authors.affiliation: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: Dave
authors.initials: D
authors.lastname: Stuart
authors.affiliation: Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Fry
authors.affiliation: The Pirbright Institute, Woking, Surrey, GU24 0NF, UK.
authors.forename: Bryan
authors.initials: B
authors.lastname: Charleston
authors.affiliation: The Pirbright Institute, Woking, Surrey, GU24 0NF, UK.
authors.forename: Julian
authors.initials: J
authors.lastname: Seago
mesh_terms: D000818:Animals; D002213:Capsid / Q000276:immunology; D036022:Capsid Proteins / Q000276:immunology*; D002460:Cell Line; D006224:Cricetinae; D005536:Foot-and-Mouth Disease / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics* / Q000276:immunology*; D016679:Genome, Viral / Q000235:genetics; D006041:Goats / Q000821:virology; D006358:Hot Temperature; D016133:Polymerase Chain Reaction; D065288:Serogroup; D014611:Vaccination; D015164:Vaccines, Inactivated / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D036022:Capsid Proteins; D015164:Vaccines, Inactivated; D014765:Viral Vaccines