title: An immunosensor based on antibody binding fragments attached to gold nanoparticles for the detection of peptides derived from avian influenza hemagglutinin H5.
abstract: This paper concerns the development of an immunosensor for detection of peptides derived from avian influenza hemagglutinin H5. Its preparation consists of successive gold electrode modification steps: (i) modification with 1,6-hexanedithiol and gold colloidal nanoparticles; (ii) immobilization of antibody-binding fragments (Fab') of anti-hemagglutinin H5 monoclonal antibodies Mab 6-9-1 via S-Au covalent bonds; and (iii) covering the remaining free space on the electrode surfaces with bovine serum albumin. The interactions between Fab' fragments and hemagglutinin (HA) variants have been explored with electrochemical impedance spectroscopy (EIS) in the presence of [Fe(CN)6](3-/4-) as an electroactive marker. The immunosensor was able to recognize three different His-tagged variants of recombinant hemagglutinin from H5N1 viruses: H1 subunit (17-340 residues) of A/swan/Poland/305-135V08/2006, the long HA (17-530 residues) A/Bar-headed Goose/Qinghai/12/2005 and H1 subunit (1-345 residues) of A/Vietnam/1194/2004. The strongest response has been observed for the long variant with detection limit of 2.2 pg/mL and dynamic range from 4.0 to 20.0 pg/mL.
authors.affiliation: Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland. u.jarocka@pan.olsztyn.pl.
authors.forename: Urszula
authors.initials: U
authors.lastname: Jarocka
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland. rozasawicka@ibb.waw.pl.
authors.forename: Róża
authors.initials: R
authors.lastname: Sawicka
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland. annag@ibb.waw.pl.
authors.forename: Anna
authors.initials: A
authors.lastname: Góra-Sochacka
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland. asirko@ibb.waw.pl.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Sirko
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland. wlodzimierz.zagorski@ibb.waw.pl.
authors.forename: Włodzimierz
authors.initials: W
authors.lastname: Zagórski-Ostoja
authors.affiliation: Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland. j.radecki@pan.olsztyn.pl.
authors.forename: Jerzy
authors.initials: J
authors.lastname: Radecki
authors.affiliation: Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland. h.radecka@pan.olsztyn.pl.
authors.forename: Hanna
authors.initials: H
authors.lastname: Radecka
mesh_terms: D000914:Antibodies, Viral / Q000276:immunology*; D015374:Biosensing Techniques / Q000295:instrumentation*; D058266:Dielectric Spectroscopy / Q000295:instrumentation; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold / Q000737:chemistry*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000032:analysis* / Q000276:immunology; D007118:Immunoassay / Q000295:instrumentation*; D007128:Immunoglobulin Fragments / Q000276:immunology; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology*; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D036103:Nanotechnology / Q000295:instrumentation; D010455:Peptides / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007128:Immunoglobulin Fragments; D010455:Peptides; D006046:Gold