title: Error-prone pcr-based mutagenesis strategy for rapidly generating high-yield influenza vaccine candidates.
abstract: Vaccination is the primary strategy for the prevention and control of influenza outbreaks. However, the manufacture of influenza vaccine requires a high-yield seed strain, and the conventional methods for generating such strains are time consuming. In this study, we developed a novel method to rapidly generate high-yield candidate vaccine strains by integrating error-prone PCR, site-directed mutagenesis strategies, and reverse genetics. We used this method to generate seed strains for the influenza A(H1N1)pdm09 virus and produced six high-yield candidate strains. We used a mouse model to assess the efficacy of two of the six candidate strains as a vaccine seed virus: both strains provided complete protection in mice against lethal challenge, thus validating our method. Results confirmed that the efficacy of these candidate vaccine seed strains was not affected by the yield-optimization procedure.
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, MS, USA.
authors.forename: Jianqiang
authors.initials: J
authors.lastname: Ye
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, MS, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Wen
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, MS, USA.
authors.forename: Yifei
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, MS, USA.
authors.forename: Nan
authors.initials: N
authors.lastname: Zhao
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, MS, USA.
authors.forename: Liping
authors.initials: L
authors.lastname: Long
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, MS, USA.
authors.forename: Hailiang
authors.initials: H
authors.lastname: Sun
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, MS, USA.
authors.forename: Jialiang
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University, MS, USA.
authors.forename: Jim
authors.initials: J
authors.lastname: Cooley
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, MS, USA.
authors.forename: G
authors.initials: G
authors.lastname: Todd Pharr
authors.affiliation: Department of Infectious Diseases, St. Jude Children׳s Research Hospital, Memphis, TN, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Webby
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, MS, USA. Electronic address: wan@cvm.msstate.edu.
authors.forename: Xiu-Feng
authors.initials: XF
authors.lastname: Wan
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D008807:Mice, Inbred BALB C; D016296:Mutagenesis; D009976:Orthomyxoviridae Infections / Q000517:prevention & control; D016133:Polymerase Chain Reaction / Q000379:methods*; D059386:Reverse Genetics; D016019:Survival Analysis; D013678:Technology, Pharmaceutical / Q000379:methods*
chemical_list: D007252:Influenza Vaccines
keywords: Error-prone PCR; High-yield strain; Influenza A virus; Vaccine; Vaccine production