Journal
BIOSENSORS & BIOELECTRONICS
Volume 55, Issue -, Pages 301-306Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2013.12.030
Keywords
Glassy carbon electrodes; Detection of antibody; Avian influenza virus; Immunosensor; Electrochemical impedance spectroscopy
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Funding
- Innovative Economy Program [WNDPOIG.01.01.02-00-007/08]
- Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Olsztyn, Poland
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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 ED/NHS coupling reaction, (iii) covering with anti-His IgG monoclonal antibody, (iv) immobilization of the recombinant His-tagged hemagglutinin (His(6)-H-5 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-H-5 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 2.1 pg/mL, the quantification limit (LOQ) of 63 pg/mL and a dynamic range from 4 pg/mL to 20 pg/mL. In addition, this analytical device was applied for detection of antibodies against His(6)-H-5 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 x 10(7)-fold. The sensitivity of immunosensor was about four orders of magnitude much better than ELISA. (C) 2013 Elsevier B.V. All rights reserved.
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