Rapid and specific influenza virus detection by functionalized magnetic nanoparticles and mass spectrometry
Journal
Journal of Nanobiotechnology
Journal Volume
9
Date Issued
2011
Author(s)
Abstract
Background: 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.Results: 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).Conclusions: 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 3EID 50per 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. ? 2011 Chou et al; licensee BioMed Central Ltd.
Subjects
Gel electrophoresis; Hemagglutinin; Influenza; Magnetic nanoparticles; Mass spectrometry; Viruses
SDGs
Other Subjects
Antiviral therapy; Cross-reactivity; Functionalized; Gel electrophoresis; Hemagglutinin; High sensitivity; High specificity; Influenza; Influenza A virus; Influenza virus; LC-MS/MS; Liquid chromatography-tandem mass spectrometry; Magnetic nanoparticles; MALDI TOF MS; MALDI-MS; Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry; Non-specific binding; Peptide sequences; Polyacrylamide gel electrophoresis; Polymerase chain reaction techniques; SDS-PAGE; Sensitive method; Sodium dodecyl sulfate; Assays; Chemical detection; Computer viruses; Desorption; Diagnosis; Diffusers (optical); Electrophoresis; Ethylene; Ethylene glycol; Gels; Iron oxides; Liquid chromatography; Magnetic separation; Mass spectrometers; Mass spectrometry; Monoclonal antibodies; Nanoparticles; Polymerase chain reaction; Probes; Sodium; Viruses; Visualization; Nanomagnetics; ethylene glycol; Influenza virus hemagglutinin; monoclonal antibody; superparamagnetic iron oxide nanoparticle; virus antibody; ferric ion; ferric oxide; magnetite nanoparticle; amino acid sequence; article; Influenza virus A H5N2; liquid chromatography; matrix assisted laser desorption ionization time of flight mass spectrometry; nonhuman; polyacrylamide gel electrophoresis; tandem mass spectrometry; virus detection; chemistry; high performance liquid chromatography; human; immunology; influenza; Influenza virus A H5N1; isolation and purification; mass spectrometry; methodology; sensitivity and specificity; tandem mass spectrometry; Influenza A virus; Orthomyxoviridae; Amino Acid Sequence; Antibodies, Viral; Chromatography, High Pressure Liquid; Electrophoresis, Polyacrylamide Gel; Ferric Compounds; Hemagglutinin Glycoproteins, Influenza Virus; Humans; Influenza A Virus, H5N1 Subtype; Influenza A Virus, H5N2 Subtype; Influenza, Human; Magnetite Nanoparticles; Sensitivity and Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tandem Mass Spectrometry
Type
journal article