Electrochemical immunosensor for detection of antibodies against influenza A virus H5N1 in hen serum
2014
Jarocka, Urszula | Sawicka, Róża | Góra-Sochacka, Anna | Sirko, Agnieszka | Zagórski-Ostoja, Włodzimierz | Radecki, Jerzy | Radecka, Hanna
This paper describes the development of an immunosensor for detection of anti-hemagglutinin antibodies. Its preparation consists of successive modification steps of glassy carbon electrodes: (i) creation of COOH groups, (ii) covalent immobilization of protein A with EDC/NHS coupling reaction, (iii) covering with anti-His IgG monoclonal antibody, (iv) immobilization of the recombinant His-tagged hemagglutinin (His6-H5 HA), (v) filling free space with BSA. The interactions between two variants of recombinant HA (short and long) from highly pathogenic avian influenza virus H5N1 and the anti-H5 HA monoclonal antibody (Mab 6-9-1) have been explored with electrochemical impedance spectroscopy (EIS). The impedimetric immunosensor displayed a very good detection limit (LOD) of 2.1pg/mL, the quantification limit (LOQ) of 6.3pg/mL and a dynamic range from 4pg/mL to 20pg/mL. In addition, this analytical device was applied for detection of antibodies against His6-H5 HA in serum of vaccinated hen using serial 10-fold dilutions of serum. The immunosensor proposed was able to detect antibody in hen serum diluted up to 7×107-fold. The sensitivity of immunosensor was about four orders of magnitude much better than ELISA.
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