title: Direct electrochemical detection of PB1-F2 protein of influenza A virus in infected cells.
abstract: Influenza virus represents a major concern of human health and animal production. PB1-F2 is a small proapoptotic protein supposed to contribute to the virulence of influenza A virus (IAV). However, the molecular mechanism of action of PB1-F2 is still unclear.PB1-F2 expression and behavior during the viral cycle is difficult to follow with classical biochemical methods. In this work we have developed an electrochemical biosensor based on immuno-detection system for quantification of PB1-F2 protein in infected cell. The electrochemical immunosensor was based on conducting copolypyrrole integrating ferrocenyl group as redox marker for enhancing signal detection. A specific anti-PB1-F2 monoclonal antibody was immobilized on the copolypyrrole layer via biotin-streptavidin system. We demonstrate that this electrochemical system sensitively detect purified recombinant PB1-F2 over a wide range of concentrations from 5 nM to 1.5 µM. The high sensor sensitivity allowed the detection of PB1-F2 in lysates of infected cells confirming that PB1-F2 is expressed in early stages of viral cycle. The immunosensor developed shows enhanced performances for the evaluation of PB1-F2 protein concentration in biological samples and could be applied for studying of PB1-F2 during influenza virus infection.
authors.affiliation: CNRS UMR-8182, Institut de Chimie Moléculaire et de Matériaux d'Orsay, Equipe de Chimie Bioorganique et Bioinorganique, Bâtiment 420, 91405 Orsay, France.
authors.forename: Anna
authors.initials: A
authors.lastname: Miodek
authors.affiliation: CNRS UMR-8182, Institut de Chimie Moléculaire et de Matériaux d'Orsay, Equipe de Chimie Bioorganique et Bioinorganique, Bâtiment 420, 91405 Orsay, France.
authors.forename: Helene
authors.initials: H
authors.lastname: Sauriat-Dorizon
authors.affiliation: INRA, Unité de Virologie et Immunologie Moléculaires, UR892, Domaine de Vilvert, F-78350, Cntre de Jouy-en-Josas, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Chevalier
authors.affiliation: INRA, Unité de Virologie et Immunologie Moléculaires, UR892, Domaine de Vilvert, F-78350, Cntre de Jouy-en-Josas, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Delmas
authors.affiliation: INRA, Unité de Virologie et Immunologie Moléculaires, UR892, Domaine de Vilvert, F-78350, Cntre de Jouy-en-Josas, France. Electronic address: jasmina.vidic@jouy.inra.fr.
authors.forename: Jasmina
authors.initials: J
authors.lastname: Vidic
authors.affiliation: CNRS UMR-8182, Institut de Chimie Moléculaire et de Matériaux d'Orsay, Equipe de Chimie Bioorganique et Bioinorganique, Bâtiment 420, 91405 Orsay, France. Electronic address: hafsa.korri-youssoufi@u-psud.fr.
authors.forename: Hafsa
authors.initials: H
authors.lastname: Korri-Youssoufi
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D002460:Cell Line; D055664:Electrochemical Techniques / Q000379:methods*; D004847:Epithelial Cells / Q000821:virology; D005296:Ferrous Compounds / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000379:methods; D009980:Influenza A virus / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D000075163:Metallocenes; D008958:Models, Molecular; D011108:Polymers / Q000737:chemistry; D011758:Pyrroles / Q000737:chemistry; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000032:analysis*
chemical_list: D005296:Ferrous Compounds; D000075163:Metallocenes; C442816:PB1-F2 protein, Influenza A virus; D011108:Polymers; D011758:Pyrroles; D014764:Viral Proteins; C067635:polypyrrole; C004998:ferrocene
keywords: Biosensor; Electrochemical; Ferrocene; Immunosensor; Influenza virus; Polypyrrole