Rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometry
<p>Abstract</p> <p>Background</p> <p>The 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...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2011-11-01
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| Series: | Journal of Nanobiotechnology |
| Subjects: | |
| Online Access: | http://www.jnanobiotechnology.com/content/9/1/52 |
| _version_ | 1811305293702758400 |
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| author | Chou Tzu-Chi Hsu Wei Wang Ching-Ho Chen Yu-Ju Fang Jim-Min |
| author_facet | Chou Tzu-Chi Hsu Wei Wang Ching-Ho Chen Yu-Ju Fang Jim-Min |
| author_sort | Chou Tzu-Chi |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>The 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.</p> <p>Results</p> <p>Iron 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).</p> <p>Conclusions</p> <p>The 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 (~10<sup>3 </sup>EID<sub>50 </sub>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.</p> |
| first_indexed | 2024-04-13T08:22:52Z |
| format | Article |
| id | doaj.art-d9750aa0a8d44e249db927a323088303 |
| institution | Directory Open Access Journal |
| issn | 1477-3155 |
| language | English |
| last_indexed | 2024-04-13T08:22:52Z |
| publishDate | 2011-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Nanobiotechnology |
| spelling | doaj.art-d9750aa0a8d44e249db927a3230883032022-12-22T02:54:35ZengBMCJournal of Nanobiotechnology1477-31552011-11-01915210.1186/1477-3155-9-52Rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometryChou Tzu-ChiHsu WeiWang Ching-HoChen Yu-JuFang Jim-Min<p>Abstract</p> <p>Background</p> <p>The 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.</p> <p>Results</p> <p>Iron 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).</p> <p>Conclusions</p> <p>The 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 (~10<sup>3 </sup>EID<sub>50 </sub>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.</p>http://www.jnanobiotechnology.com/content/9/1/52VirusesInfluenzaHemagglutininMagnetic nanoparticlesMass spectrometryGel electrophoresis |
| spellingShingle | Chou Tzu-Chi Hsu Wei Wang Ching-Ho Chen Yu-Ju Fang Jim-Min Rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometry Journal of Nanobiotechnology Viruses Influenza Hemagglutinin Magnetic nanoparticles Mass spectrometry Gel electrophoresis |
| title | Rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometry |
| title_full | Rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometry |
| title_fullStr | Rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometry |
| title_full_unstemmed | Rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometry |
| title_short | Rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometry |
| title_sort | rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometry |
| topic | Viruses Influenza Hemagglutinin Magnetic nanoparticles Mass spectrometry Gel electrophoresis |
| url | http://www.jnanobiotechnology.com/content/9/1/52 |
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