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Sensors 2014, 14(9), 15714-15728; doi:10.3390/s140915714

An Immunosensor Based on Antibody Binding Fragments Attached to Gold Nanoparticles for the Detection of Peptides Derived from Avian Influenza Hemagglutinin H5

1
Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland
2
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland
*
Author to whom correspondence should be addressed.
Received: 19 May 2014 / Revised: 10 July 2014 / Accepted: 11 August 2014 / Published: 25 August 2014
(This article belongs to the Special Issue Immunosensors 2014)
View Full-Text   |   Download PDF [864 KB, uploaded 25 August 2014]   |  

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. View Full-Text
Keywords: avian influenza virus; Fab’ fragments; gold electrodes; electrochemical immunosensor; electrochemical impedance spectroscopy avian influenza virus; Fab’ fragments; gold electrodes; electrochemical immunosensor; electrochemical impedance spectroscopy
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Jarocka, U.; Sawicka, R.; Góra-Sochacka, A.; Sirko, A.; Zagórski-Ostoja, W.; Radecki, J.; Radecka, H. An Immunosensor Based on Antibody Binding Fragments Attached to Gold Nanoparticles for the Detection of Peptides Derived from Avian Influenza Hemagglutinin H5. Sensors 2014, 14, 15714-15728.

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