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

Sensors

Urszula Jarocka¹  Ró៊ Sawicka²  Anna Góra-Sochacka²  Agnieszka Sirko²  Włodzimierz Zagórski-Ostoja²  Jerzy Radecki¹ 

[1] Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland; E-Mails:;Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland; E-Mails:

关键词: avian influenza virus;    Fab' fragments;    gold electrodes;    electrochemical immunosensor;    electrochemical impedance spectroscopy;   

DOI  :  10.3390/s140915714

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.