期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 407, 期 25, 页码 7807-7814出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-015-8949-y
关键词
Dipyrromethene-Cu(II) layer; Histidine-tagged antigen immobilization; Detection of antibodies in sera; Avian influenza virus
资金
- National Centre for Research and Development (NCBiR) [PBS2/A7/14/2014, 679/N-BELGIA/2010/0]
- COST Action [CM10005]
- Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland
- University of Leuven
- FWO-Vlaanderen
- Ministerie voor Wetenschapsbeleid
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 x 10(6), which is about 200 times better than the enzyme-linked immunosorbent assay.
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