Rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometry

Journal of Nanobiotechnology

Research

Tzu-Chi Chou¹  Yu-Ju Chen²  Jim-Min Fang³  Ching-Ho Wang⁴  Wei Hsu⁵ 

[1] Department of Chemistry, National Taiwan University, 106, Taipei, Taiwan;Department of Chemistry, National Taiwan University, 106, Taipei, Taiwan;Institute of Chemistry, Academia Sinica, 115, Taipei, Taiwan;Department of Chemistry, National Central University, 320, Jhong-Li, Taiwan;Department of Chemistry, National Taiwan University, 106, Taipei, Taiwan;The Genomics Research Center, Academia Sinica, 115, Taipei, Taiwan;Department of Veterinary Medicine, National Taiwan University, 106, Taipei, Taiwan;Institute of Chemistry, Academia Sinica, 115, Taipei, Taiwan;Department of Chemistry, National Central University, 320, Jhong-Li, Taiwan;

关键词: Viruses;    Influenza;    Hemagglutinin;    Magnetic nanoparticles;    Mass spectrometry;    Gel electrophoresis;   

DOI  :  10.1186/1477-3155-9-52

 received in 2011-07-06, accepted in 2011-11-16,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundThe timely and accurate diagnosis of specific influenza virus strains is crucial to effective prophylaxis, vaccine preparation and early antiviral therapy. The detection of influenza A viruses is mainly accomplished using polymerase chain reaction (PCR) techniques or antibody-based assays. In conjugation with the immunoassay utilizing monoclonal antibody, mass spectrometry is an alternative to identify proteins derived from a target influenza virus. Taking advantage of the large surface area-to-volume ratio, antibody-conjugated magnetic nanoparticles can act as an effective probe to extract influenza virus for sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and on-bead mass spectrometric analysis.ResultsIron oxide magnetic nanoparticles (MNP) were functionalized with H5N2 viral antibodies targeting the hemagglutinin protein and capped with methoxy-terminated ethylene glycol to suppress nonspecific binding. The antibody-conjugated MNPs possessed a high specificity to H5N2 virus without cross-reactivity with recombinant H5N1 viruses. The unambiguous identification of the captured hemagglutinin on magnetic nanoparticles was realized by SDS-PAGE visualization and peptide sequence identification using liquid chromatography-tandem mass spectrometry (LC-MS/MS).ConclusionsThe assay combining efficient magnetic separation and MALDI-MS readout offers a rapid and sensitive method for virus screening. Direct on-MNP detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) provided high sensitivity (~103 EID50 per mL) and a timely diagnosis within one hour. The magnetic nanoparticles encapsulated with monoclonal antibodies could be used as a specific probe to distinguish different subtypes of influenza.

【 授权许可】

Unknown   
© Chou et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.