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• W3049025057 abstract "Influenza A/H5N1 viruses caused several panzootic outbreaks in the past. These viruses of avian origin have acquired a zoonotic potential by genetic changes and can cross species barriers including humans. There is an urgent need of reliable broad spectrum H5N1 diagnostics. The aim of this work is to identify H5 specific antibodies applicable in a subtype specific diagnostic detection and discrimination system for influenza viruses.Two single chain (sc)Fv antibody gene phage libraries were generated from chickens vaccinated and challenged with avian H5N1 influenza viruses from phylogenetically distant clades. For antibody selection two additional recombinant H5 antigens were applied. Twelve unique anti-H5 scFvs were isolated expressed in mammalian cells. All of them showed binding to H5 hemagglutinin proteins in ELISA and immunoblot assays without cross-reactivity with unrelated proteins. The majority of selected binders detected a large variety of H5 viruses in immunoperoxidase monolayer assays (IPMA) and showed no cross-reactivity with other influenza viruses of closely related HA-subtypes. For one antibody very broad binding specificity for H5 antigens from different strains and for two antibodies neutralizing activity was demonstrated. Epitope mapping revealed a six amino acid short epitope highly conserved among H5 influenza viruses. In H5 sandwich ELISA and membrane based immunodot blot assays the proof of principle of a rapid diagnostic assay,was demonstrated. Antibodies specifically stained influenza viruses of an H5 strain in electron microscopy.Thus, a successful immunization and selection strategy comprising four different H5 sources leading to several H5 specific antibodies and elucidating a highly conserved linear epitope specific for H5 influenza viruses has been demonstrated. Isolated antibodies and the highly conserved linear epitope in the H5 HA1 domain can be used in different applications e.g. influenza diagnostics, therapy and vaccination." @default.
• W3049025057 created "2020-08-21" @default.
• W3049025057 creator A5000626092 @default.
• W3049025057 date "2012-10-17" @default.
• W3049025057 modified "2023-09-26" @default.
• W3049025057 title "Generation of Recombinant Designer Antibodies Using Phage Display for a Rapid and Subtype Specific InfluenzaDetection System" @default.
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