A biosensor based on electroactive dipyrromethene-Cu(II) layer deposited onto gold electrodes for the detection of antibodies against avian influenza virus type H5N1 in hen sera.

Abstract

This paper describes the development of a biosensor for the detection of anti-hemagglutinin antibodies against the influenza virus hemagglutinin. The steps of biosensor fabrications are as follows: (i) creation of a mixed layer containing the thiol derivative of dipyrromethene and 4-mercapto-1-butanol, (ii) complexation of Cu(II) ions, (iii) oriented immobilization of the recombinant histidine-tagged hemagglutinin, and (iv) filling free spaces with bovine serum albumin. The interactions between recombinants hemagglutinin from the highly pathogenic avian influenza virus type H5N1 and anti-hemagglutinin H5 monoclonal antibodies were explored with Osteryoung square-wave voltammetry. The biosensor displayed a good detection limit of 2.4 pg/mL, quantification limit of 7.2 pg/mL, and dynamic range from 4.0 to 100.0 pg/mL in buffer. In addition, this analytical device was applied for the detection of antibodies in hen sera from individuals vaccinated and non-vaccinated against the avian influenza virus type H5N1. The limit of detection for the assay was the dilution of sera 1: 7 × 10(6), which is about 200 times better than the enzyme-linked immunosorbent assay.

• Authors: Urszula Jarocka, Róża Sawicka, Anna Stachyra, Anna Góra-Sochacka, Agnieszka Sirko, Włodzimierz Zagórski-Ostoja, Violetta Sączyńska, Anna Porębska, Wim Dehaen, Jerzy Radecki, Hanna Radecka
• Journal: Analytical and bioanalytical chemistry (Anal Bioanal Chem) Vol. 407 Issue 25 Pg. 7807-14 (Oct 2015) ISSN: 1618-2650 [Electronic] Germany
• PMID: 26297459 (Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Viral
• Hemagglutinin Glycoproteins, Influenza Virus
• dipyrromethene
• hemagglutinin, avian influenza A virus
• Gold
• Porphobilinogen
• Copper

Topics

• Animals
• Antibodies, Viral (blood, immunology)
• Biosensing Techniques (instrumentation)
• Chickens (blood, immunology, virology)
• Copper (chemistry)
• Electrochemical Techniques (instrumentation)
• Electrodes
• Female
• Gold (chemistry)
• Hemagglutinin Glycoproteins, Influenza Virus (immunology)
• Immunoassay (instrumentation)
• Influenza A Virus, H5N1 Subtype (immunology, isolation & purification)
• Influenza in Birds (blood, diagnosis, immunology, virology)
• Limit of Detection
• Porphobilinogen (analogs & derivatives, chemistry)