Journal: Influenza and Other Respiratory Viruses

Article Title: SERS‐based immunomagnetic bead for rapid detection of H5N1 influenza virus

doi: 10.1111/irv.13114

Figure Lengend Snippet: Schematic of the IMBSIs@Ag‐SERS method for H5N1 influenza virus detection.

Article Snippet: Then, 1 μg/ml, 0.5 μg/ml, 0.33 μg/ml, 0.25 μg/ml and 0.1 μg/ml of biotinylated rabbit anti‐H5N1 antibody (Sino Biological Inc., Beijing, China) were added to the MB‐SA and incubated for 1 h at room temperature with gentle rotation in the tube.

Techniques:

Journal: Influenza and Other Respiratory Viruses

Article Title: SERS‐based immunomagnetic bead for rapid detection of H5N1 influenza virus

doi: 10.1111/irv.13114

Figure Lengend Snippet: Detection of H5N1 influenza virus by IMBSIs@Ag‐SERS method. (A) Effect of SERS enhancement in situ reduction of nano‐silver. (B) SERS of H5N1 influenza virus captured by the primary antibodies in different concentrations. (C) TEM of naked magnetic beads with streptavidin in situ reduction of nano‐silver. (D) TEM of immunomagnetic beads sandwich immunocomplexes (IMBSIs) in situ reduction of nano‐silver.

Article Snippet: Then, 1 μg/ml, 0.5 μg/ml, 0.33 μg/ml, 0.25 μg/ml and 0.1 μg/ml of biotinylated rabbit anti‐H5N1 antibody (Sino Biological Inc., Beijing, China) were added to the MB‐SA and incubated for 1 h at room temperature with gentle rotation in the tube.

Techniques: In Situ, Magnetic Beads

Journal: Influenza and Other Respiratory Viruses

Article Title: SERS‐based immunomagnetic bead for rapid detection of H5N1 influenza virus

doi: 10.1111/irv.13114

Figure Lengend Snippet: Excellent repeatability and specificity of SERS detection in H5N1 influenza virus. (A) The IMBSIs@Ag SERS showed a consistent Raman intensity at 1053 cm −1 for nine times. (B) The IMBSIs@Ag SERS no characteristic peak at 1053 cm −1 replacing H5N1 with H1N1, H5N6 or H9N2.

Article Snippet: Then, 1 μg/ml, 0.5 μg/ml, 0.33 μg/ml, 0.25 μg/ml and 0.1 μg/ml of biotinylated rabbit anti‐H5N1 antibody (Sino Biological Inc., Beijing, China) were added to the MB‐SA and incubated for 1 h at room temperature with gentle rotation in the tube.

Techniques:

Journal: Influenza and Other Respiratory Viruses

Article Title: SERS‐based immunomagnetic bead for rapid detection of H5N1 influenza virus

doi: 10.1111/irv.13114

Figure Lengend Snippet: The susceptibility of IMBSIs@Ag for detection of the H5N1 influenza virus. (A) IMBSIs@Ag Raman intensity of the H5N1 influenza virus was 10‐fold diluted from 5.0 × 10 6 to 5.0 × 10 −7 TCID 50 /ml. (B) Linear relationship between Raman intensity and H5N1 influenza virus concentration. (C) IMBSIs@Ag SERS identification of H5N1 influenza virus from chicken embryos.

Article Snippet: Then, 1 μg/ml, 0.5 μg/ml, 0.33 μg/ml, 0.25 μg/ml and 0.1 μg/ml of biotinylated rabbit anti‐H5N1 antibody (Sino Biological Inc., Beijing, China) were added to the MB‐SA and incubated for 1 h at room temperature with gentle rotation in the tube.

Techniques: Concentration 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: 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