Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Production of H5N1 avian influenza triple H5N1/NA-HA-M1 VLPs. ( A ) Schematic representation of the tricistronic expression cassette. ( B ) Western blotting analysis of NA-HA-M1 VLPs purified on a 10%–60% sucrose gradient. Antibodies used for the detection of HA, NA and M1 proteins: mouse monoclonal anti-HA H5N1 antibodies, mouse monoclonal anti-M1 influenza antibodies and rabbit polyclonal anti-avian influenza A neuraminidase antibody. ( C ) Schematic representation of the predicted structure of triple H5N1/NA-HA-M1 VLPs. Structural proteins are color-coded according to the scheme of the expression cassette: NA—yellow, HA—blue, M1—green.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Expressing, Western Blot, Purification

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Characterization of H5N1 avian influenza virus triple H5N1/NA-HA-M1 VLPs. ( A ) Transmission electron microscopy of triple H5N1/NA-HA-M1 VLPs and influenza A/H5N2 virus. Scale bar = 1 µm is shown in the right bottom corner of the images. ( B ) Hemagglutination assay. The AIV A/ostrich/Denmark/725/96 (H5N2) was used as a positive control. The HA titer was determined as the reciprocal of the highest dilution with HA activity. Each dot represents HA titers obtained in each experiment. The bars represent the median values of obtained HA titers. ( C ) Neuraminidase activity assay. The AIV A/Ck/Scot/59 (H5N1) was used as a positive control. For each assay, the mean value from three independent experiments performed is presented. The mean A560 values and standard deviations are shown on the y-axis.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Virus, Transmission Assay, Electron Microscopy, Hemagglutination Assay, Positive Control, Activity Assay

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Schematic timeline of immunization of the broiler hens with triple H5N1/NA-HA-M1 VLPs. Red droplets represent the days of blood collection.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques:

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: End-point titration of chicken sera after immunization with triple H5N1/NA-HA-M1 VLPs. ELISA plates were coated with reference antigen inactivated H5N1 virus. Serial dilutions of chicken A/H5N1/HPAI polyclonal antibodies served as a positive control. Chicken serum from day 0 and control chicken serum served as a background. The dilution factor of the pooled sera is shown on the x-axis. For each ELISA, the mean value from three independent experiments performed is presented. The mean A450 values and standard deviations are shown on the y-axis.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Titration, Enzyme-linked Immunosorbent Assay, Virus, Positive Control, Control

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Dynamics of anti-H5N1 IgY level in sera of immunized chickens. Kinetics of antibody titer in chickens ( n = 5) following prime and boost vaccinations with triple H5N1/NA-HA-M1 VLPs were measured via ELISA test. The median (thick line) is shown with the interquartile 25% and 75% range (narrow lines). The day of the serum collection is shown on the x-axis. The A450 values are shown on the y-axis. Statistical analysis was performed using the nonparametric Kruskal–Wallis test ( p = 0.05) and Benjamini, Krieger and Yekutieli multiple comparison test ( p = 0.05). Statistical differences were detected between days 0–41 and 0–34 ( p = 0.014) and shown on the graph as a star symbol.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: HI titer of pooled chicken sera collected after immunization with triple H5N1/NA-HA-M1 VLPs. H5N1 A/Ck/Scot/59 and H5N2 A/Ost/Den/72420/96 antibodies were used as a positive control. Sera from chickens vaccinated with PBS/ICF mixture served as a negative control. HI assay was performed in triplicates.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Positive Control, Negative Control, HI Assay

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Construction and characterization of HA-stalk universal influenza antigen from homologous H5N1 HPAI virus strain. ( A ) Schematic representation of the full-length HA (top panel) and HA-stalk constructs (bottom panel). To obtain the HA-stalk construct, a glycine linker was added between cysteines in positions C52 and C277, replacing the head region of the HA1 domain. ( B ) Schematic representation of the predicted structure of HA monomer and HA-stalk monomer. ( C ) Expression of H5N1 HA-stalk in insect cells was confirmed by IPMA with anti-H5N1 monoclonal antibodies. Full-length HA from the H5N1 strain was used as a positive control. Cells infected with the wild type baculovirus were used as a negative control. Images were taken at ×10 magnification. ( D ) Reactivity of the HA-stalk antigen and full-length HA from H5N1 in the IPMA with broadly neutralizing universal FI6 human antibodies. HA-stalk and full-length HA from the H5N1 strain was detected in transfected insect cells. Images were taken at ×20 magnification.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Virus, Construct, Expressing, Positive Control, Infection, Negative Control, Transfection

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Expression of HA-stalk antigens from the 1 and 2 HA groups in mammalian cells. HEK293 cells were transfected with HA-stalk H1N1, H5N1 and H7N9 constructs. Protein expression was detected using different monoclonal antibodies specific for H1N1, H5N1 and H7N9 influenza strains. Non-transfected cells were used as a negative control. Images were taken at ×40 magnification.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Expressing, Transfection, Construct, Negative Control

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Cross-reactivity of chicken sera obtained after vaccination with triple H5N1/NA-HA-M1 VLPs with the HA-stalk antigens from the 1 and 2 HA groups. Detection of HA-stalk from H1N1, H5N1 and H7N9 strains was performed on transfected HEK293 cells. Sera from unvaccinated hens were used as a background. Non-transfected cells were used as a negative control. Images were taken at ×40 magnification.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Transfection, Negative Control

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Alignment of amino acid sequences coding LAH regions from H5N1, pH1N1, H7N9 and H3N2 HA protein. ( A ) Alignment view with consensus sequence where the highest similarity is shown as a green colour. ( B ) Matrix showing the percentage of sequence identity between sequences.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Sequencing

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Cross-reactivity of chicken sera obtained after vaccination with triple H5N1/NA-HA-M1 VLPs with the LAH peptide from H3 from 2 HA group. The antibody titer in chickens ( n = 5) before (gray) and after (black) vaccination with triple H5N1/NA-HA-M1 VLPs was measured via the peptide ELISA test. The mean OD values and standard deviations are shown on the y-axis. The tested chickens are shown on the x-axis. Statistical analysis was performed using a nonparametric Wilcoxon test ( p = 0.05) for paired groups.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Peptide ELISA

Journal: Sensors (Basel, Switzerland)

Article Title: A Homogenous Fluorescence Quenching Based Assay for Specific and Sensitive Detection of Influenza Virus A Hemagglutinin Antigen

doi: 10.3390/s150408852

Figure Lengend Snippet: Cross-reactivity of two assays with human influenza virus H1N1 HA and avian influenza virus H5N1-HA. The mixtures, prepared with 6G-QDs/mAb-H1-Au NPs and with 3GQDs/mAb-H5-Au NPs, were tested using HAs derived from A/New Caledonia/20/1999 and A/Viet Nam/1194/2004 (120 nM), respectively. The control experiment was carried out using BSA (150 μM), (N = 3).

Article Snippet: Recombinant influenza A virus H1N1 (A/New Caledonia/20/1999) HA, recombinant influenza A virus H5N1 (A/Viet Nam/1194/2004) HA, mouse monoclonal antibody to influenza A virus H1N1 HA (mAb-H1-HA), and mouse monoclonal antibody to influenza A virus H5N1 HA (mAb-H5-HA) were purchased from Sino Biological Inc. (Beijing, China).

Techniques: Derivative Assay

Journal: Sensors (Basel, Switzerland)

Article Title: A Homogenous Fluorescence Quenching Based Assay for Specific and Sensitive Detection of Influenza Virus A Hemagglutinin Antigen

doi: 10.3390/s150408852

Figure Lengend Snippet: Detection of human influenza virus H1N1-HA and avian H5N1-HA by 6G-QDs/mAb- H1-Au NPs mixture and 3G-QDs/mAb-H5-Au NPs mixture, in PBS ( a ) and in human sera spiked PBS ( b ), respectively. (N = 3).

Article Snippet: Recombinant influenza A virus H1N1 (A/New Caledonia/20/1999) HA, recombinant influenza A virus H5N1 (A/Viet Nam/1194/2004) HA, mouse monoclonal antibody to influenza A virus H1N1 HA (mAb-H1-HA), and mouse monoclonal antibody to influenza A virus H5N1 HA (mAb-H5-HA) were purchased from Sino Biological Inc. (Beijing, China).

Techniques:

Journal: Sensors (Basel, Switzerland)

Article Title: A Homogenous Fluorescence Quenching Based Assay for Specific and Sensitive Detection of Influenza Virus A Hemagglutinin Antigen

doi: 10.3390/s150408852

Figure Lengend Snippet: Comparison of the sensitivity of the developed biosensors for the detection of influenza virus.

Article Snippet: Recombinant influenza A virus H1N1 (A/New Caledonia/20/1999) HA, recombinant influenza A virus H5N1 (A/Viet Nam/1194/2004) HA, mouse monoclonal antibody to influenza A virus H1N1 HA (mAb-H1-HA), and mouse monoclonal antibody to influenza A virus H5N1 HA (mAb-H5-HA) were purchased from Sino Biological Inc. (Beijing, China).

Techniques: Derivative Assay

Journal: Communications Biology

Article Title: A risk marker of tribasic hemagglutinin cleavage site in influenza A (H9N2) virus

doi: 10.1038/s42003-020-01589-7

Figure Lengend Snippet: a Maximum likelihood (ML) phylogeny of the H9N2 influenza virus HA gene. The HA gene sequences from all available H9N2 viruses were downloaded from GISAID’s EpiFlu Database for phylogenetic analysis. The ML tree was inferred with the RAxML software under the GTRGAMMA model with 1,000 bootstraps. References H9N2 viruses from each HACS motif are denoted by different colors. The distribution of different HACS motif of influenza A (H9N2) viruses from GISAID’s EpiFlu and GenBank Database. The branch length is scaled according to the numbers of substitutions per site (subs/site). b represents the distribution of the amino acids of HACS except for China, and c represents the amino acids of HACS in China. Different colored bars represent multiple HACS motifs of influenza A(H9N2) viruses. Amino acids frequency of all available HACS of influenza A(H9N2) viruses. Frequencies of each position, from P6 to P4’ of the HACS are illustrated using WebLogo 3.4 ( http://weblogo.threeplusone.com/ ). d represents China, and e represents all countries except for China.

Article Snippet: Polyclonal rabbit anti-HA antibody from A/Hong Kong/1073/99 H9N2 expressed in baculovirus (1:2000 dilution for 2 h at room temperature; Sino Biological Inc., Beijing, China) was used to detect HA.

Techniques: Software

Journal: Communications Biology

Article Title: A risk marker of tribasic hemagglutinin cleavage site in influenza A (H9N2) virus

doi: 10.1038/s42003-020-01589-7

Figure Lengend Snippet: Locations of human infections with H9N2 viruses are indicated by the blue cartoon figure. In the pie charts, the size represents the number of H9N2 isolates, and the colors represents different motifs of HA cleavage sites (HACS). The size of the dotted pie chart does not represent the number of H9N2 isolates in China owing to a large quantity of strains. Data are available from the GISAID’s EpiFlu Database, GenBank Database, the World Health Organization, the World Organisation for Animal Health, and the National Health and Family Planning Commission of the People’s Republic of China. The map was designed using ArcGIS Desktop 10.4 software ( http://www.esri.com/software/arcgis/arcgis-for-desktop/ ).

Article Snippet: Polyclonal rabbit anti-HA antibody from A/Hong Kong/1073/99 H9N2 expressed in baculovirus (1:2000 dilution for 2 h at room temperature; Sino Biological Inc., Beijing, China) was used to detect HA.

Techniques: Software

Journal: Communications Biology

Article Title: A risk marker of tribasic hemagglutinin cleavage site in influenza A (H9N2) virus

doi: 10.1038/s42003-020-01589-7

Figure Lengend Snippet: Growth curves after inoculation of each virus at a multiplicity of infection (MOI) of 0.001 into ( a ) MDCK cells and at an MOI of 0.001 into ( b ) CEF cells. Each point on the curve is the mean ± standard deviation from three independent experiments. The independent-samples t test was used for analysis. c Thermo stability of the influenza A(H9N2) viruses. The influenza A(H9N2) viruses were incubated for 1, 2, 3, 4, and 5 h at 50 °C. The titers of heat-treated recombination viruses were determined by TCID 50 assay in MDCK cells. Western blots of CEF ( d ) and A549 ( e ) cell cultures inoculated with five influenza A(H9N2) virus at an MOI of 0.01 in the presence of 1 μ g/ml trypsin for 24 h. f Syncytia formation induced by five influenza A(H9N2) viruses in Vero cells. Cultures of Vero cells were inoculated with H9N2 viruses at MOI of 1 for 16 h, followed by incubation with 10 μ g/ml of TPCK-trypsin for 15 min to cleave expressed HA0. To induce syncytia formation, Vero cells were incubated for 5 min with fusion buffers of pH values ranging from 5.0 to 5.8, with 0.2 unit increments, and were visualized 3 h later. Representative images at pH values of 5.0 to 5.8 derived from three replicates are shown. Five groups of ten 5-week-old SPF chickens were infected intranasally with 10 6 EID 50 /200 μL of each virus. g Viral titers in the lungs of three inoculated chickens. h Viral titers in the duodenum of three inoculated chickens. i Viral titers in the kidney of three inoculated chickens. j Viral titers in the brain of three inoculated chickens. k Antibody titers of inoculated and exposed chickens at 14 dpi. An HI titer ≥1:16 was considered seroconversion. Five groups of three female BALB/c mice were intranasally inoculated with 10 6 EID 50 /50 μL of the H9N2 viruses. Three mice from each group were euthanized at 4 dpi. Viral titers in the mouse lungs ( l ) were shown. The dashed lines represent the detection limit. The independent-samples t test was used for analysis.

Article Snippet: Polyclonal rabbit anti-HA antibody from A/Hong Kong/1073/99 H9N2 expressed in baculovirus (1:2000 dilution for 2 h at room temperature; Sino Biological Inc., Beijing, China) was used to detect HA.

Techniques: Infection, Standard Deviation, Incubation, Western Blot, Derivative Assay