title: 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: Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748, Olsztyn, Poland.
authors: Urszula
authors: U
authors: Jarocka
authors: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106, Warsaw, Poland.
authors: Róża
authors: R
authors: Sawicka
authors: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106, Warsaw, Poland.
authors: Anna
authors: A
authors: Stachyra
authors: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106, Warsaw, Poland.
authors: Anna
authors: A
authors: Góra-Sochacka
authors: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106, Warsaw, Poland.
authors: Agnieszka
authors: A
authors: Sirko
authors: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106, Warsaw, Poland.
authors: Włodzimierz
authors: W
authors: Zagórski-Ostoja
authors: Institute of Biotechnology and Antibiotics, Starościńska 5, 02-516, Warsaw, Poland.
authors: Violetta
authors: V
authors: Sączyńska
authors: Institute of Biotechnology and Antibiotics, Starościńska 5, 02-516, Warsaw, Poland.
authors: Anna
authors: A
authors: Porębska
authors: Department of Chemistry, University of Leuven, Celestijnenlaan 200F, 3001, Leuven, Belgium.
authors: Wim
authors: W
authors: Dehaen
authors: Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748, Olsztyn, Poland.
authors: Jerzy
authors: J
authors: Radecki
authors: Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748, Olsztyn, Poland. h.radecka@pan.olsztyn.pl.
authors: Hanna
authors: H
authors: Radecka
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation*; D002645:Chickens / Q000097:blood* / Q000276:immunology / Q000821:virology; D003300:Copper / Q000737:chemistry; D055664:Electrochemical Techniques / Q000295:instrumentation; D004566:Electrodes; D005260:Female; D006046:Gold / Q000737:chemistry; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D007118:Immunoassay / Q000295:instrumentation; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology* / Q000302:isolation & purification; D005585:Influenza in Birds / Q000097:blood* / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D057230:Limit of Detection; D011162:Porphobilinogen / Q000031:analogs & derivatives / Q000737:chemistry
chemical_list: D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; C507544:dipyrromethene; C109963:hemagglutinin, avian influenza A virus; D006046:Gold; D011162:Porphobilinogen; D003300:Copper
keywords: Avian influenza virus; Detection of antibodies in sera; Dipyrromethene-Cu(II) layer; Histidine-tagged antigen immobilization