Table 1

Examples of influenza virus-like particle production
Influenza viral protein Influenza strain DNA vector system Cell expression platform Purification Characterization quantification Ref
HA,NA, M1, M2 A/Udorn/72 (H3N2) rBV Sf9 insect cells Partial purification and concentration by ultracentrifugation/Iodixanol and sucrose gradient ultracentrifugation EM/WB/SDS Page [19]
HA (H2/H7/H5) NA, M1 A/Indonesia/05/2005(H5N1) rBV Sf9 insect cells Sucrose step gradient ultracentrifugation EM/Quantitative-WB/SDS Page/SRID assay/Hemagglutination assay [20]
A/Swine/Missouri/4296424/2006 (H2N3)
A/New York/107/2003 (H7N2)
A/Viet Nam/1203/2004 (H5N1)
HA, NA, M1 A/HongKong/1073/99 (H9N2) rBV Sf9 insect cells Partial purification and concentration by ultracentrifugation/Sucrose gradient ultracentrifugation EM/Quantitative WB/SDS Page/Neuraminidase activity/Hemagglutination assay [13]
HA, M1 A/Puerto Rico/8/1934 (H1N1) rBV Sf9 insect cells Partial purification and concentration by ultracentrifugation/Sucrose gradient ultracentrifugation EM/WB/SDS Page/Hemagglutination assay [21]
A/WSN/33
HA, NA, M1 A/Fujian/411/2002 (H3N2) rBV Sf9 insect cells Sucrose gradient ultracentrifugation/Ion-exchange chromatography EM/WB/SDS Page/TBA Neuraminidase assay/Hemagglutination assay/SRID assay [22,23]
A/Indonesia/5/2005 (H5N1)
HA, M1 HA: A/California/04/2009 (H1N1) rBV Sf9 & BTI-TN5B1-4 insect cells Partial purification and concentration by ultracentrifugation/Sucrose gradient ultracentrifugation EM/Quantitative WB/SDS Page/Hemagglutination assay [16]
M1: A/Udorn/307/1972 (H3N2)
NA A/Cambodia/JP52a/2005 (H5N1) pCDNA plasmid HEK 293T – HeLa – A549 mammalian cells Partial purification and concentration by sucrose cushion ultracentrifugation/Sucrose gradient ultracentrifugation EM/Quantitative WB/SDS Page/NA Star Neuraminidase assay [24]
HA, NA, NP, M1, M2, NS2, PB1, PB2, PA A/Udorn/72 (H3N2) pCAGGS plasmid HEK 293T & HeLa mammalian cells Partial purification and concentration by sucrose cushion ultracentrifugation/Sucrose gradient ultracentrifugation EM/Quantitative WB/SDS Page/VLP infectivity assay (transfer of GFP-expressing pseudogene) [25]
HA, NA, M2 A/Chicken/FPV/Rostock/1934 (H7N1) MLV GagPol HEK 293T & TE671 human cells Partial purification and concentration by sucrose cushion ultracentrifugation Quantitative WB/SDS Page [26,27]
A/Thailand/KAN-1/04 (H5N1)
HA, NA, M1 A/Vietnam/1203/2004 (H5N1) rMVA vector Vero monkey cells Partial Purification and concentration by filtration with 100kDa molecular weight cut off/Sucrose gradient ultracentrifugation EM/WB/SDS Page/Hemagglutination assay [28]
A/Puerto Rico/8/1934 (H1N1)
HA, NA, M1 A/Puerto Rico/8/1934 (H1N1) rBV HEK 293T human cells Partial purification and concentration by sucrose cushion ultracentrifugation/Sucrose gradient ultracentrifugation/Precipitation by ultracentrifugation BCA Total protein assay/WB/SDS Page/Hemagglutination assay [17]
HA, M1 A/Indonesia/5/05 (H5N1) rAgrobacterium N. benthamiana plant cells Centrifugation/Affinity chromatography EM/WB/SDS Page/Total protein analysis Lipid analysis/Hemagglutination assay [29]
A/New Caledonia/20/99 (H1N1)
HA, NA, M1, M2 A/Taiwian/083/2006 (H3N2) pC14 plasmid Vero monkey cells Partial purification and concentration by sucrose cushion ultracentrifugation/Sucrose gradient ultracentrifugation EM/WB/SDS Page/Total protein analysis/Hemagglutination assay/Dynamic Light Scattering/LC-MS/MS [12,30]
A/Hanoi/30408/2005 (H5N1)

Associated expression system, quantification and purification applied (rBV, Recombinant baculovirus; MLV, Murine Leukemia Virus).

Thompson et al.

Thompson et al. Virology Journal 2013 10:141   doi:10.1186/1743-422X-10-141

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