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Biosens Bioelectron. 2013 Dec 21;55C:301-306. doi: 10.1016/j.bios.2013.12.030. [Epub ahead of print]
Electrochemical immunosensor for detection of antibodies against influenza A virus H5N1 in hen serum.
Jarocka U1, Sawicka R2, G?ra-Sochacka A2, Sirko A2, Zag?rski-Ostoja W2, Radecki J1, Radecka H3.
Author information
Abstract
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.
Copyright ? 2013 Elsevier B.V. All rights reserved.
KEYWORDS:
Avian influenza virus, Detection of antibody, Electrochemical impedance spectroscopy, Glassy carbon electrodes, Immunosensor
PMID:
24412426
[PubMed - as supplied by publisher]
Electrochemical immunosensor for detection of antibodies against influenza A virus H5N1 in hen serum.
Jarocka U1, Sawicka R2, G?ra-Sochacka A2, Sirko A2, Zag?rski-Ostoja W2, Radecki J1, Radecka H3.
Author information
Abstract
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.
Copyright ? 2013 Elsevier B.V. All rights reserved.
KEYWORDS:
Avian influenza virus, Detection of antibody, Electrochemical impedance spectroscopy, Glassy carbon electrodes, Immunosensor
PMID:
24412426
[PubMed - as supplied by publisher]
