influenza a h1n1 hemagglutinin ha antibody mouse mab  (Sino Biological)


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    Name:
    Influenza A H1N1 A California 07 2009 Hemagglutinin HA Antibody Mouse MAb
    Description:
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified recombinant Influenza A H1N1 A California 07 2009 Hemagglutinin HA Catalog 11085 V08H ACP44189 1 Met1 Gln529 The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography
    Catalog Number:
    11085-MM01
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    H1N1
    Applications:
    ELISA
    Immunogen:
    Recombinant Influenza A H1N1 (A/California/07/2009) Hemagglutinin / HA Protein (Catalog#11085-V08H)
    Antibody Type:
    MAb
    Host:
    Mouse
    Isotype:
    Mouse IgG1
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    Structured Review

    Sino Biological influenza a h1n1 hemagglutinin ha antibody mouse mab
    Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1. As a first step, <t>influenza</t> A (H1N1) virus HA, NA, and NP peptide T cell recognition was tested with spleen cells from Pandemrix-vaccinated HLA-DQ6.2 mice, restimulated with pools of 5 peptides each (15-mers). Pools that stimulated IFN -γ or IL-2 expression were broken up, and single peptides tested either with spleen cells from additional Pandemrix-vaccinated HLA-DQ6.2 mice, or PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. As a second step, recognition of single peptides was then tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls. As a third step, influenza A (H1N1) T cell peptides that showed increased stimulation of IFN - γ or IL-2 secretion in patients vs. controls were validated and mapped. Cross-reactive T cell self-epitopes were predicted by BLAST against human proteome, and recognition tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls.
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified recombinant Influenza A H1N1 A California 07 2009 Hemagglutinin HA Catalog 11085 V08H ACP44189 1 Met1 Gln529 The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography
    https://www.bioz.com/result/influenza a h1n1 hemagglutinin ha antibody mouse mab/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    influenza a h1n1 hemagglutinin ha antibody mouse mab - by Bioz Stars, 2021-09
    94/100 stars

    Images

    1) Product Images from "Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1"

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1

    Journal: Nature Communications

    doi: 10.1038/s41467-021-22637-8

    Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1. As a first step, influenza A (H1N1) virus HA, NA, and NP peptide T cell recognition was tested with spleen cells from Pandemrix-vaccinated HLA-DQ6.2 mice, restimulated with pools of 5 peptides each (15-mers). Pools that stimulated IFN -γ or IL-2 expression were broken up, and single peptides tested either with spleen cells from additional Pandemrix-vaccinated HLA-DQ6.2 mice, or PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. As a second step, recognition of single peptides was then tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls. As a third step, influenza A (H1N1) T cell peptides that showed increased stimulation of IFN - γ or IL-2 secretion in patients vs. controls were validated and mapped. Cross-reactive T cell self-epitopes were predicted by BLAST against human proteome, and recognition tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls.
    Figure Legend Snippet: Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1. As a first step, influenza A (H1N1) virus HA, NA, and NP peptide T cell recognition was tested with spleen cells from Pandemrix-vaccinated HLA-DQ6.2 mice, restimulated with pools of 5 peptides each (15-mers). Pools that stimulated IFN -γ or IL-2 expression were broken up, and single peptides tested either with spleen cells from additional Pandemrix-vaccinated HLA-DQ6.2 mice, or PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. As a second step, recognition of single peptides was then tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls. As a third step, influenza A (H1N1) T cell peptides that showed increased stimulation of IFN - γ or IL-2 secretion in patients vs. controls were validated and mapped. Cross-reactive T cell self-epitopes were predicted by BLAST against human proteome, and recognition tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls.

    Techniques Used: Mouse Assay, Expressing

    Influenza A H1N1 virus T-cell epitope screen in HLA-DQ6.2 mice, and Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. Spleen cells from Pandemrix-immunized HLA-DQ6.2 mice were stimulated in culture with pools of five overlapping 15-mer peptides each, covering a hemagglutinin (HA), b neuraminidase (NA) or c nucleoprotein (NP) from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus used in Pandemrix (pooling cells from 2 × 2 mice, n = 2). Recombinant hemagglutinin (rHA) and nucleoprotein (rNP; A/Puerto Rico/8/34 corresponding to the Pandemrix vaccine strain) were used as positive controls. d – f PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals (sleep clinic patients without a diagnosis of NT1) were stimulated in culture with single 15-mer peptides, derived from the same vaccine virus strain ( n = 1–2). The expression of IFN-γ or IL-2 was measured by FMIA (protein) or RT-qPCR (mRNA). Results are expressed as ratios between cytokine concentrations (lines representing means) or relative gene expressions (dots representing single values, bars representing means) measured in peptide-stimulated and negative control samples (stimulation index). An asterisk (*) indicates a pool or single peptide that was selected for further testing.
    Figure Legend Snippet: Influenza A H1N1 virus T-cell epitope screen in HLA-DQ6.2 mice, and Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. Spleen cells from Pandemrix-immunized HLA-DQ6.2 mice were stimulated in culture with pools of five overlapping 15-mer peptides each, covering a hemagglutinin (HA), b neuraminidase (NA) or c nucleoprotein (NP) from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus used in Pandemrix (pooling cells from 2 × 2 mice, n = 2). Recombinant hemagglutinin (rHA) and nucleoprotein (rNP; A/Puerto Rico/8/34 corresponding to the Pandemrix vaccine strain) were used as positive controls. d – f PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals (sleep clinic patients without a diagnosis of NT1) were stimulated in culture with single 15-mer peptides, derived from the same vaccine virus strain ( n = 1–2). The expression of IFN-γ or IL-2 was measured by FMIA (protein) or RT-qPCR (mRNA). Results are expressed as ratios between cytokine concentrations (lines representing means) or relative gene expressions (dots representing single values, bars representing means) measured in peptide-stimulated and negative control samples (stimulation index). An asterisk (*) indicates a pool or single peptide that was selected for further testing.

    Techniques Used: Mouse Assay, Recombinant, Derivative Assay, Expressing, Quantitative RT-PCR, Negative Control

    Mapping of identified influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. a , d PBMC from pediatric Pandemrix-associated NT1 patients (NT1; validation cohort) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with overlapping 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus neuraminidase (NA) or nucleoprotein (NP), as indicated. b , c , e , f PBMC from NT1 patients (invariably HLA-DQB1*0602 positive; discovery and validation cohorts combined; HLA-DQB1*0602 homozygous NT1 patients marked with red dots) or HLA-DQB1*0602 positive (C/DQ6+) or negative (C/DQ6−) healthy controls were stimulated with single NA- or NP-derived peptides. The secretion of IFN-γ was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.
    Figure Legend Snippet: Mapping of identified influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. a , d PBMC from pediatric Pandemrix-associated NT1 patients (NT1; validation cohort) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with overlapping 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus neuraminidase (NA) or nucleoprotein (NP), as indicated. b , c , e , f PBMC from NT1 patients (invariably HLA-DQB1*0602 positive; discovery and validation cohorts combined; HLA-DQB1*0602 homozygous NT1 patients marked with red dots) or HLA-DQB1*0602 positive (C/DQ6+) or negative (C/DQ6−) healthy controls were stimulated with single NA- or NP-derived peptides. The secretion of IFN-γ was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.

    Techniques Used: Derivative Assay, Negative Control

    Identification of influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. PBMC from pediatric Pandemrix-associated NT1 patients (NT1) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with single 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus hemagglutinin (HA), neuraminidase (NA) or nucleoprotein (NP) (discovery cohort). Recombinant neuraminidase (rNA) and nucleoprotein (rNP) were used as positive controls. The secretion of IFN-γ ( a , c , e ) or IL-2 ( b , d , f ) was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.
    Figure Legend Snippet: Identification of influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. PBMC from pediatric Pandemrix-associated NT1 patients (NT1) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with single 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus hemagglutinin (HA), neuraminidase (NA) or nucleoprotein (NP) (discovery cohort). Recombinant neuraminidase (rNA) and nucleoprotein (rNP) were used as positive controls. The secretion of IFN-γ ( a , c , e ) or IL-2 ( b , d , f ) was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.

    Techniques Used: Recombinant, Negative Control

    2) Product Images from "Antigenic Differences between AS03 Adjuvanted Influenza A (H1N1) Pandemic Vaccines: Implications for Pandemrix-Associated Narcolepsy Risk"

    Article Title: Antigenic Differences between AS03 Adjuvanted Influenza A (H1N1) Pandemic Vaccines: Implications for Pandemrix-Associated Narcolepsy Risk

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0114361

    Antibody levels to HA as antibody titers determined with the hemagglutination inhibition (HI) test. Antibody titers against influenza A/California/07/2009 vaccine strain (A), epidemic A/Finland/814/01 (H1N1) virus strain (B) and influenza A/Finland/715/00 (H3N2) virus strain (C) in plasma samples from 47 children with narcolepsy and in 57 healthy vaccinated children.
    Figure Legend Snippet: Antibody levels to HA as antibody titers determined with the hemagglutination inhibition (HI) test. Antibody titers against influenza A/California/07/2009 vaccine strain (A), epidemic A/Finland/814/01 (H1N1) virus strain (B) and influenza A/Finland/715/00 (H3N2) virus strain (C) in plasma samples from 47 children with narcolepsy and in 57 healthy vaccinated children.

    Techniques Used: HI Assay

    Related Articles

    other:

    Article Title: Aptamer-Antibody Complementation On Multiwalled Carbon Nanotube-Gold Transduced Dielectrode Surfaces To Detect Pandemic Swine Influenza Virus
    Article Snippet: HA of A/California/07/2009 (H1N1) and A/Panama/2007/1999 (H3N2) and Anti-HA A/California/07/2009 antibodies were procured from Sino Biological (Beijing, China).

    Article Title: Pandemic H1N1 influenza A directly induces a robust and acute inflammatory gene signature in primary human bronchial epithelial cells downstream of membrane fusion.
    Article Snippet: Protein was expressed from DNA sequences encoding the extracellular regions of A/Brisbane/59/ 2007 (H1N1) or A/California/07/2009 (H1N1) HA.

    Recombinant:

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1
    Article Snippet: .. Peptides and recombinant proteins15-mer peptides covering hemagglutinin (GenBank entry ACP41953.1) and neuraminidase (YP_009118627.1) of influenza (A/California/07/2009 (H1N1)), and nucleoprotein (ADE29096.1) of influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) with 12 amino acid overlap were produced. ..

    Produced:

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1
    Article Snippet: .. Peptides and recombinant proteins15-mer peptides covering hemagglutinin (GenBank entry ACP41953.1) and neuraminidase (YP_009118627.1) of influenza (A/California/07/2009 (H1N1)), and nucleoprotein (ADE29096.1) of influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) with 12 amino acid overlap were produced. ..

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    Sino Biological influenza a virus hemagglutinin ha antibody rabbit mab
    Effect of λ-CGN on <t>influenza</t> A infection in vivo . BALB/c mice were mock-infected (black) or intranasally infected with maPR8 at 5 MLD50 (red). As test groups, the virus was preincubated at room temperature for 30 min with λ-CGN at a lower dose (1 mg/kg/d, bright green) or a higher dose (5 mg/kg/d, dark green), followed by intranasal administration. Control mice received oral OSV-P twice a day (10 mg/kg/d) at 8-h intervals, starting at 4 h before viral infection (blue). Body weight (A) and mortality (B) of mice were measured every day from Days 0 to 14 post-infection. Data are expressed as the mean ± SEM from five mice
    Influenza A Virus Hemagglutinin Ha Antibody Rabbit Mab, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/influenza a virus hemagglutinin ha antibody rabbit mab/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    influenza a virus hemagglutinin ha antibody rabbit mab - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    94
    Sino Biological influenza a h1n1 hemagglutinin ha antibody mouse mab
    Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1. As a first step, <t>influenza</t> A (H1N1) virus HA, NA, and NP peptide T cell recognition was tested with spleen cells from Pandemrix-vaccinated HLA-DQ6.2 mice, restimulated with pools of 5 peptides each (15-mers). Pools that stimulated IFN -γ or IL-2 expression were broken up, and single peptides tested either with spleen cells from additional Pandemrix-vaccinated HLA-DQ6.2 mice, or PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. As a second step, recognition of single peptides was then tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls. As a third step, influenza A (H1N1) T cell peptides that showed increased stimulation of IFN - γ or IL-2 secretion in patients vs. controls were validated and mapped. Cross-reactive T cell self-epitopes were predicted by BLAST against human proteome, and recognition tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls.
    Influenza A H1n1 Hemagglutinin Ha Antibody Mouse Mab, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/influenza a h1n1 hemagglutinin ha antibody mouse mab/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    influenza a h1n1 hemagglutinin ha antibody mouse mab - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    93
    Sino Biological mouse monoclonal antibodies against ha
    High-resolution MES-PAGE analysis of Pandemrix and Arepandrix vaccines under reducing and non-reducing conditions. Pandemrix H1N1 antigen suspension run under reducing conditions (A) with a major band including a mixture of <t>HA</t> and NP, and under non-reducing conditions (B) with HA shown in the upper area and NP at its original place identical to RF position of the reducing gel (A). Recombinant NP from H1N1/A/Puerto Rico 1934 (Sino Biologicals Inc., China) alone run under reducing conditions (C). HA stained with <t>monoclonal</t> <t>mouse</t> <t>anti-HA</t> antibody (Sino Biologicals Inc., China) using western blotting of the separated proteins of Pandemrix H1N1 antigen suspension under non-reducing conditions (D) and under reducing conditions (E). Arepanrix H1N1 antigen suspension run under reducing conditions (F) with a major band including a mixture of HA and NP (F), and run under non-reducing conditions (G) with HA shown in the upper area and NP at its original place identical to RF position of the reducing gel (F).
    Mouse Monoclonal Antibodies Against Ha, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal antibodies against ha/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal antibodies against ha - by Bioz Stars, 2021-09
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    Image Search Results


    Effect of λ-CGN on influenza A infection in vivo . BALB/c mice were mock-infected (black) or intranasally infected with maPR8 at 5 MLD50 (red). As test groups, the virus was preincubated at room temperature for 30 min with λ-CGN at a lower dose (1 mg/kg/d, bright green) or a higher dose (5 mg/kg/d, dark green), followed by intranasal administration. Control mice received oral OSV-P twice a day (10 mg/kg/d) at 8-h intervals, starting at 4 h before viral infection (blue). Body weight (A) and mortality (B) of mice were measured every day from Days 0 to 14 post-infection. Data are expressed as the mean ± SEM from five mice

    Journal: bioRxiv

    Article Title: Antiviral activity of lambda-carrageenan against influenza viruses in mice and severe acute respiratory syndrome coronavirus 2 in vitro

    doi: 10.1101/2020.08.23.255364

    Figure Lengend Snippet: Effect of λ-CGN on influenza A infection in vivo . BALB/c mice were mock-infected (black) or intranasally infected with maPR8 at 5 MLD50 (red). As test groups, the virus was preincubated at room temperature for 30 min with λ-CGN at a lower dose (1 mg/kg/d, bright green) or a higher dose (5 mg/kg/d, dark green), followed by intranasal administration. Control mice received oral OSV-P twice a day (10 mg/kg/d) at 8-h intervals, starting at 4 h before viral infection (blue). Body weight (A) and mortality (B) of mice were measured every day from Days 0 to 14 post-infection. Data are expressed as the mean ± SEM from five mice

    Article Snippet: Viral NP and HA proteins were detected using mouse anti-NP (catalog no. 11675-MM03; Sino Biological, Beijing, China) and rabbit anti-HA2 (catalog no. 86001-RM01; Sino Biological) antibodies, respectively, according to our previous report .

    Techniques: Infection, In Vivo, Mouse Assay

    Effect of λ-CGN on the intracellular entry of influenza A virus. MDCK cells were infected with PR8 (MOI, 5) and subsequently mock-treated or treated either with λ-CGN or with p-KG03 at a concentration of 10 μg/ml. At 4 h p.i. in the absence of CHX (A) or at 2.5 h in the presence of 10 μg/ml CHX (B), viral NP was detected with an anti-NP antibody and an Alex Fluor 488-conjugated goat anti-mouse secondary antibody (green). Cell nuclei were counterstained with DAPI (blue). Original magnification, 400×.

    Journal: bioRxiv

    Article Title: Antiviral activity of lambda-carrageenan against influenza viruses in mice and severe acute respiratory syndrome coronavirus 2 in vitro

    doi: 10.1101/2020.08.23.255364

    Figure Lengend Snippet: Effect of λ-CGN on the intracellular entry of influenza A virus. MDCK cells were infected with PR8 (MOI, 5) and subsequently mock-treated or treated either with λ-CGN or with p-KG03 at a concentration of 10 μg/ml. At 4 h p.i. in the absence of CHX (A) or at 2.5 h in the presence of 10 μg/ml CHX (B), viral NP was detected with an anti-NP antibody and an Alex Fluor 488-conjugated goat anti-mouse secondary antibody (green). Cell nuclei were counterstained with DAPI (blue). Original magnification, 400×.

    Article Snippet: Viral NP and HA proteins were detected using mouse anti-NP (catalog no. 11675-MM03; Sino Biological, Beijing, China) and rabbit anti-HA2 (catalog no. 86001-RM01; Sino Biological) antibodies, respectively, according to our previous report .

    Techniques: Infection, Concentration Assay

    Antiviral effect of λ-CGN against influenza A virus in vivo. BALB/c mice (6–7 weeks old female) were mock-infected (black) or intranasally infected with maPR8 at 5 MLD 50 (red). As test groups, the virus was preincubated at room temperature for 30 min with λ-CGN at a lower dose (1 mg/kg/d, purple) or a higher dose (5 mg/kg/d, green), followed by intranasal administration. Control mice received OSV-P orally twice a day (10 mg/kg/d) at 8-h intervals, starting at 4 h before viral infection (blue). Body weight ( A ) and mortality ( B ) of mice were measured every day from days 0 to 14 post-infection. Data are expressed as the mean ± S.D. from five mice. Survival statistics were calculated by Log-rank (Mantel–Cox) test. ** P

    Journal: Scientific Reports

    Article Title: Antiviral activity of lambda-carrageenan against influenza viruses and severe acute respiratory syndrome coronavirus 2

    doi: 10.1038/s41598-020-80896-9

    Figure Lengend Snippet: Antiviral effect of λ-CGN against influenza A virus in vivo. BALB/c mice (6–7 weeks old female) were mock-infected (black) or intranasally infected with maPR8 at 5 MLD 50 (red). As test groups, the virus was preincubated at room temperature for 30 min with λ-CGN at a lower dose (1 mg/kg/d, purple) or a higher dose (5 mg/kg/d, green), followed by intranasal administration. Control mice received OSV-P orally twice a day (10 mg/kg/d) at 8-h intervals, starting at 4 h before viral infection (blue). Body weight ( A ) and mortality ( B ) of mice were measured every day from days 0 to 14 post-infection. Data are expressed as the mean ± S.D. from five mice. Survival statistics were calculated by Log-rank (Mantel–Cox) test. ** P

    Article Snippet: Influenza viral NP and HA proteins were detected using mouse anti-NP (Cat. No., 11675-MM03; Sino Biological, Beijing, China) and rabbit anti-HA2 (Cat. No., 86001-RM01; Sino Biological) antibodies, respectively, according to our previous report .

    Techniques: In Vivo, Mouse Assay, Infection

    Effect of λ-CGN on the influenza A virus entry. ( A ) HA inhibition assay. Two-fold serially diluted PR8 (from 2 3 to 2 11 ) in PBS was incubated with an equal volume of PBS or twofold increasing concentrations of λ-CGN for 20 min. HA titer in each combination was determined at 30 min after addition of 0.5% chicken RBC. HA titers are marked on the right side of the panel. ( B , C ) Confocal microscopy. MDCK cells were infected with PR8 (MOI, 5) in the absence (Mock) or presence of either λ-CGN or p-KG03 at a concentration of 10 μg/ml. At 4 h post-infection in the absence of CHX ( B ) or at 2.5 h in the presence of 10 μg/ml CHX ( C ), viral NP was detected with an anti-NP antibody and an Alex Fluor 488-conjugated goat anti-mouse secondary antibody (green). Cell nuclei were counterstained with DAPI (blue). Original magnification, 400×. The images were analyzed using ZEN blue software 3.1 ( www.zeiss.com ).

    Journal: Scientific Reports

    Article Title: Antiviral activity of lambda-carrageenan against influenza viruses and severe acute respiratory syndrome coronavirus 2

    doi: 10.1038/s41598-020-80896-9

    Figure Lengend Snippet: Effect of λ-CGN on the influenza A virus entry. ( A ) HA inhibition assay. Two-fold serially diluted PR8 (from 2 3 to 2 11 ) in PBS was incubated with an equal volume of PBS or twofold increasing concentrations of λ-CGN for 20 min. HA titer in each combination was determined at 30 min after addition of 0.5% chicken RBC. HA titers are marked on the right side of the panel. ( B , C ) Confocal microscopy. MDCK cells were infected with PR8 (MOI, 5) in the absence (Mock) or presence of either λ-CGN or p-KG03 at a concentration of 10 μg/ml. At 4 h post-infection in the absence of CHX ( B ) or at 2.5 h in the presence of 10 μg/ml CHX ( C ), viral NP was detected with an anti-NP antibody and an Alex Fluor 488-conjugated goat anti-mouse secondary antibody (green). Cell nuclei were counterstained with DAPI (blue). Original magnification, 400×. The images were analyzed using ZEN blue software 3.1 ( www.zeiss.com ).

    Article Snippet: Influenza viral NP and HA proteins were detected using mouse anti-NP (Cat. No., 11675-MM03; Sino Biological, Beijing, China) and rabbit anti-HA2 (Cat. No., 86001-RM01; Sino Biological) antibodies, respectively, according to our previous report .

    Techniques: Inhibition, Incubation, Confocal Microscopy, Infection, Concentration Assay, Software

    Inhibition of infection by influenza A virus (H1N1) HA/NA- or SARS-CoV-2 spike-pseudotyped viruses. Lentiviral pseudotypes bearing influenza A virus (H1N1) HA and NA proteins (black bars) or SARS-CoV-2 spike protein (gray bars) were prepared, in which a firefly luciferase-expressing plasmid was incorporated. They were preincubated with mock or increasing concentrations of λ-CGN for 2 h at 37 °C and then infected into Vero E6 cells. On day 2, relative firefly luciferase activity (RLU) was determined by fixing the mock-treated sample at 100%. Data are expressed as the mean ± S.D. of three independent experiments. **** P

    Journal: Scientific Reports

    Article Title: Antiviral activity of lambda-carrageenan against influenza viruses and severe acute respiratory syndrome coronavirus 2

    doi: 10.1038/s41598-020-80896-9

    Figure Lengend Snippet: Inhibition of infection by influenza A virus (H1N1) HA/NA- or SARS-CoV-2 spike-pseudotyped viruses. Lentiviral pseudotypes bearing influenza A virus (H1N1) HA and NA proteins (black bars) or SARS-CoV-2 spike protein (gray bars) were prepared, in which a firefly luciferase-expressing plasmid was incorporated. They were preincubated with mock or increasing concentrations of λ-CGN for 2 h at 37 °C and then infected into Vero E6 cells. On day 2, relative firefly luciferase activity (RLU) was determined by fixing the mock-treated sample at 100%. Data are expressed as the mean ± S.D. of three independent experiments. **** P

    Article Snippet: Influenza viral NP and HA proteins were detected using mouse anti-NP (Cat. No., 11675-MM03; Sino Biological, Beijing, China) and rabbit anti-HA2 (Cat. No., 86001-RM01; Sino Biological) antibodies, respectively, according to our previous report .

    Techniques: Inhibition, Infection, Luciferase, Expressing, Plasmid Preparation, Activity Assay

    Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1. As a first step, influenza A (H1N1) virus HA, NA, and NP peptide T cell recognition was tested with spleen cells from Pandemrix-vaccinated HLA-DQ6.2 mice, restimulated with pools of 5 peptides each (15-mers). Pools that stimulated IFN -γ or IL-2 expression were broken up, and single peptides tested either with spleen cells from additional Pandemrix-vaccinated HLA-DQ6.2 mice, or PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. As a second step, recognition of single peptides was then tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls. As a third step, influenza A (H1N1) T cell peptides that showed increased stimulation of IFN - γ or IL-2 secretion in patients vs. controls were validated and mapped. Cross-reactive T cell self-epitopes were predicted by BLAST against human proteome, and recognition tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls.

    Journal: Nature Communications

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1

    doi: 10.1038/s41467-021-22637-8

    Figure Lengend Snippet: Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1. As a first step, influenza A (H1N1) virus HA, NA, and NP peptide T cell recognition was tested with spleen cells from Pandemrix-vaccinated HLA-DQ6.2 mice, restimulated with pools of 5 peptides each (15-mers). Pools that stimulated IFN -γ or IL-2 expression were broken up, and single peptides tested either with spleen cells from additional Pandemrix-vaccinated HLA-DQ6.2 mice, or PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. As a second step, recognition of single peptides was then tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls. As a third step, influenza A (H1N1) T cell peptides that showed increased stimulation of IFN - γ or IL-2 secretion in patients vs. controls were validated and mapped. Cross-reactive T cell self-epitopes were predicted by BLAST against human proteome, and recognition tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls.

    Article Snippet: Peptides and recombinant proteins15-mer peptides covering hemagglutinin (GenBank entry ACP41953.1) and neuraminidase (YP_009118627.1) of influenza (A/California/07/2009 (H1N1)), and nucleoprotein (ADE29096.1) of influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) with 12 amino acid overlap were produced.

    Techniques: Mouse Assay, Expressing

    Influenza A H1N1 virus T-cell epitope screen in HLA-DQ6.2 mice, and Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. Spleen cells from Pandemrix-immunized HLA-DQ6.2 mice were stimulated in culture with pools of five overlapping 15-mer peptides each, covering a hemagglutinin (HA), b neuraminidase (NA) or c nucleoprotein (NP) from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus used in Pandemrix (pooling cells from 2 × 2 mice, n = 2). Recombinant hemagglutinin (rHA) and nucleoprotein (rNP; A/Puerto Rico/8/34 corresponding to the Pandemrix vaccine strain) were used as positive controls. d – f PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals (sleep clinic patients without a diagnosis of NT1) were stimulated in culture with single 15-mer peptides, derived from the same vaccine virus strain ( n = 1–2). The expression of IFN-γ or IL-2 was measured by FMIA (protein) or RT-qPCR (mRNA). Results are expressed as ratios between cytokine concentrations (lines representing means) or relative gene expressions (dots representing single values, bars representing means) measured in peptide-stimulated and negative control samples (stimulation index). An asterisk (*) indicates a pool or single peptide that was selected for further testing.

    Journal: Nature Communications

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1

    doi: 10.1038/s41467-021-22637-8

    Figure Lengend Snippet: Influenza A H1N1 virus T-cell epitope screen in HLA-DQ6.2 mice, and Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. Spleen cells from Pandemrix-immunized HLA-DQ6.2 mice were stimulated in culture with pools of five overlapping 15-mer peptides each, covering a hemagglutinin (HA), b neuraminidase (NA) or c nucleoprotein (NP) from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus used in Pandemrix (pooling cells from 2 × 2 mice, n = 2). Recombinant hemagglutinin (rHA) and nucleoprotein (rNP; A/Puerto Rico/8/34 corresponding to the Pandemrix vaccine strain) were used as positive controls. d – f PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals (sleep clinic patients without a diagnosis of NT1) were stimulated in culture with single 15-mer peptides, derived from the same vaccine virus strain ( n = 1–2). The expression of IFN-γ or IL-2 was measured by FMIA (protein) or RT-qPCR (mRNA). Results are expressed as ratios between cytokine concentrations (lines representing means) or relative gene expressions (dots representing single values, bars representing means) measured in peptide-stimulated and negative control samples (stimulation index). An asterisk (*) indicates a pool or single peptide that was selected for further testing.

    Article Snippet: Peptides and recombinant proteins15-mer peptides covering hemagglutinin (GenBank entry ACP41953.1) and neuraminidase (YP_009118627.1) of influenza (A/California/07/2009 (H1N1)), and nucleoprotein (ADE29096.1) of influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) with 12 amino acid overlap were produced.

    Techniques: Mouse Assay, Recombinant, Derivative Assay, Expressing, Quantitative RT-PCR, Negative Control

    Mapping of identified influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. a , d PBMC from pediatric Pandemrix-associated NT1 patients (NT1; validation cohort) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with overlapping 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus neuraminidase (NA) or nucleoprotein (NP), as indicated. b , c , e , f PBMC from NT1 patients (invariably HLA-DQB1*0602 positive; discovery and validation cohorts combined; HLA-DQB1*0602 homozygous NT1 patients marked with red dots) or HLA-DQB1*0602 positive (C/DQ6+) or negative (C/DQ6−) healthy controls were stimulated with single NA- or NP-derived peptides. The secretion of IFN-γ was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.

    Journal: Nature Communications

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1

    doi: 10.1038/s41467-021-22637-8

    Figure Lengend Snippet: Mapping of identified influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. a , d PBMC from pediatric Pandemrix-associated NT1 patients (NT1; validation cohort) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with overlapping 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus neuraminidase (NA) or nucleoprotein (NP), as indicated. b , c , e , f PBMC from NT1 patients (invariably HLA-DQB1*0602 positive; discovery and validation cohorts combined; HLA-DQB1*0602 homozygous NT1 patients marked with red dots) or HLA-DQB1*0602 positive (C/DQ6+) or negative (C/DQ6−) healthy controls were stimulated with single NA- or NP-derived peptides. The secretion of IFN-γ was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.

    Article Snippet: Peptides and recombinant proteins15-mer peptides covering hemagglutinin (GenBank entry ACP41953.1) and neuraminidase (YP_009118627.1) of influenza (A/California/07/2009 (H1N1)), and nucleoprotein (ADE29096.1) of influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) with 12 amino acid overlap were produced.

    Techniques: Derivative Assay, Negative Control

    Identification of influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. PBMC from pediatric Pandemrix-associated NT1 patients (NT1) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with single 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus hemagglutinin (HA), neuraminidase (NA) or nucleoprotein (NP) (discovery cohort). Recombinant neuraminidase (rNA) and nucleoprotein (rNP) were used as positive controls. The secretion of IFN-γ ( a , c , e ) or IL-2 ( b , d , f ) was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.

    Journal: Nature Communications

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1

    doi: 10.1038/s41467-021-22637-8

    Figure Lengend Snippet: Identification of influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. PBMC from pediatric Pandemrix-associated NT1 patients (NT1) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with single 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus hemagglutinin (HA), neuraminidase (NA) or nucleoprotein (NP) (discovery cohort). Recombinant neuraminidase (rNA) and nucleoprotein (rNP) were used as positive controls. The secretion of IFN-γ ( a , c , e ) or IL-2 ( b , d , f ) was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.

    Article Snippet: Peptides and recombinant proteins15-mer peptides covering hemagglutinin (GenBank entry ACP41953.1) and neuraminidase (YP_009118627.1) of influenza (A/California/07/2009 (H1N1)), and nucleoprotein (ADE29096.1) of influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) with 12 amino acid overlap were produced.

    Techniques: Recombinant, Negative Control

    High-resolution MES-PAGE analysis of Pandemrix and Arepandrix vaccines under reducing and non-reducing conditions. Pandemrix H1N1 antigen suspension run under reducing conditions (A) with a major band including a mixture of HA and NP, and under non-reducing conditions (B) with HA shown in the upper area and NP at its original place identical to RF position of the reducing gel (A). Recombinant NP from H1N1/A/Puerto Rico 1934 (Sino Biologicals Inc., China) alone run under reducing conditions (C). HA stained with monoclonal mouse anti-HA antibody (Sino Biologicals Inc., China) using western blotting of the separated proteins of Pandemrix H1N1 antigen suspension under non-reducing conditions (D) and under reducing conditions (E). Arepanrix H1N1 antigen suspension run under reducing conditions (F) with a major band including a mixture of HA and NP (F), and run under non-reducing conditions (G) with HA shown in the upper area and NP at its original place identical to RF position of the reducing gel (F).

    Journal: PLoS ONE

    Article Title: Antigenic Differences between AS03 Adjuvanted Influenza A (H1N1) Pandemic Vaccines: Implications for Pandemrix-Associated Narcolepsy Risk

    doi: 10.1371/journal.pone.0114361

    Figure Lengend Snippet: High-resolution MES-PAGE analysis of Pandemrix and Arepandrix vaccines under reducing and non-reducing conditions. Pandemrix H1N1 antigen suspension run under reducing conditions (A) with a major band including a mixture of HA and NP, and under non-reducing conditions (B) with HA shown in the upper area and NP at its original place identical to RF position of the reducing gel (A). Recombinant NP from H1N1/A/Puerto Rico 1934 (Sino Biologicals Inc., China) alone run under reducing conditions (C). HA stained with monoclonal mouse anti-HA antibody (Sino Biologicals Inc., China) using western blotting of the separated proteins of Pandemrix H1N1 antigen suspension under non-reducing conditions (D) and under reducing conditions (E). Arepanrix H1N1 antigen suspension run under reducing conditions (F) with a major band including a mixture of HA and NP (F), and run under non-reducing conditions (G) with HA shown in the upper area and NP at its original place identical to RF position of the reducing gel (F).

    Article Snippet: Milk powder (4%) diluted in TBS-0.05% Tween20 was used for blocking, and mouse monoclonal antibodies against HA or NP (anti-HA MAb (11055-MM01) or anti- NP MAb (11675-MM03), Sino Biologicals) diluted at the concentration of 1 µg/mL in Tween- TBS was used for the identification of viral HA and NP.

    Techniques: Polyacrylamide Gel Electrophoresis, Recombinant, Staining, Western Blot

    Western-blot analysis of the presence of NP in Pandemrix and Arepanrix H1N1 antigen suspension. Pandemrix and Arepanrix H1N1 antigen suspensions were run under reducing conditions and NP stained with monoclonal mouse NP-antibody (Sino Biologicals Inc., China).

    Journal: PLoS ONE

    Article Title: Antigenic Differences between AS03 Adjuvanted Influenza A (H1N1) Pandemic Vaccines: Implications for Pandemrix-Associated Narcolepsy Risk

    doi: 10.1371/journal.pone.0114361

    Figure Lengend Snippet: Western-blot analysis of the presence of NP in Pandemrix and Arepanrix H1N1 antigen suspension. Pandemrix and Arepanrix H1N1 antigen suspensions were run under reducing conditions and NP stained with monoclonal mouse NP-antibody (Sino Biologicals Inc., China).

    Article Snippet: Milk powder (4%) diluted in TBS-0.05% Tween20 was used for blocking, and mouse monoclonal antibodies against HA or NP (anti-HA MAb (11055-MM01) or anti- NP MAb (11675-MM03), Sino Biologicals) diluted at the concentration of 1 µg/mL in Tween- TBS was used for the identification of viral HA and NP.

    Techniques: Western Blot, Staining

    Dot blot detection of viral proteins with mouse monoclonal anti-HA, anti-NA and anti-NP antibodies (Sino Biologicals Inc., China) in the density fractions isolated with gradient ultracentrifugation from Pandemrix H1N1 antigen suspension. HA and NA was detected as lipid-protein micelles in the density fraction corresponding density between 1.063–1.21 g/mL, and soluble HA, NA and NP were detected in the density fraction above 1.21 g/mL.

    Journal: PLoS ONE

    Article Title: Antigenic Differences between AS03 Adjuvanted Influenza A (H1N1) Pandemic Vaccines: Implications for Pandemrix-Associated Narcolepsy Risk

    doi: 10.1371/journal.pone.0114361

    Figure Lengend Snippet: Dot blot detection of viral proteins with mouse monoclonal anti-HA, anti-NA and anti-NP antibodies (Sino Biologicals Inc., China) in the density fractions isolated with gradient ultracentrifugation from Pandemrix H1N1 antigen suspension. HA and NA was detected as lipid-protein micelles in the density fraction corresponding density between 1.063–1.21 g/mL, and soluble HA, NA and NP were detected in the density fraction above 1.21 g/mL.

    Article Snippet: Milk powder (4%) diluted in TBS-0.05% Tween20 was used for blocking, and mouse monoclonal antibodies against HA or NP (anti-HA MAb (11055-MM01) or anti- NP MAb (11675-MM03), Sino Biologicals) diluted at the concentration of 1 µg/mL in Tween- TBS was used for the identification of viral HA and NP.

    Techniques: Dot Blot, Isolation

    A sandwich EIA for the detection of IgG-antibodies to H1N1 proteins captured from H1N1 antigen of Pandemrix with solid-phase bound antibodies to HA, NA and NP. A. Capture of H1N1 viral proteins from Pandemrix antigen in a sandwich EIA revealed that HA and NA are found in a complex in the Pandemrix H1N1 antigen, whereas NP does not complex with HA or NA. H1N1 viral proteins from Pandemrix vaccine were captured with rabbit monoclonal anti-HA, anti-NA and anti-NP antibodies bound to EIA plate and mouse monoclonal anti-HA, anti-NA and anti-NP antibodies were used as the detection antibodies. B. IgG-antibodies binding to Pandemrix derived NP were higher in children with narcolepsy (N; n = 47) than in vaccinated healthy children(C; n = 57) when studied with sandwich EIA using anti-NP antibody to capture NP from Pandemrix H1N1 antigen suspension. The children with narcolepsy (N) had higher levels of IgG-antibodies to Pandemrix derived NP than the healthy, vaccinated children without HLA DQB1*06∶02 risk allele (C/DQ6−; n = 37), whereas no difference was seen in comparison with the healthy, vaccinated children with HLA DQB1*06∶02 risk allele (C/DQ6+; n = 20).

    Journal: PLoS ONE

    Article Title: Antigenic Differences between AS03 Adjuvanted Influenza A (H1N1) Pandemic Vaccines: Implications for Pandemrix-Associated Narcolepsy Risk

    doi: 10.1371/journal.pone.0114361

    Figure Lengend Snippet: A sandwich EIA for the detection of IgG-antibodies to H1N1 proteins captured from H1N1 antigen of Pandemrix with solid-phase bound antibodies to HA, NA and NP. A. Capture of H1N1 viral proteins from Pandemrix antigen in a sandwich EIA revealed that HA and NA are found in a complex in the Pandemrix H1N1 antigen, whereas NP does not complex with HA or NA. H1N1 viral proteins from Pandemrix vaccine were captured with rabbit monoclonal anti-HA, anti-NA and anti-NP antibodies bound to EIA plate and mouse monoclonal anti-HA, anti-NA and anti-NP antibodies were used as the detection antibodies. B. IgG-antibodies binding to Pandemrix derived NP were higher in children with narcolepsy (N; n = 47) than in vaccinated healthy children(C; n = 57) when studied with sandwich EIA using anti-NP antibody to capture NP from Pandemrix H1N1 antigen suspension. The children with narcolepsy (N) had higher levels of IgG-antibodies to Pandemrix derived NP than the healthy, vaccinated children without HLA DQB1*06∶02 risk allele (C/DQ6−; n = 37), whereas no difference was seen in comparison with the healthy, vaccinated children with HLA DQB1*06∶02 risk allele (C/DQ6+; n = 20).

    Article Snippet: Milk powder (4%) diluted in TBS-0.05% Tween20 was used for blocking, and mouse monoclonal antibodies against HA or NP (anti-HA MAb (11055-MM01) or anti- NP MAb (11675-MM03), Sino Biologicals) diluted at the concentration of 1 µg/mL in Tween- TBS was used for the identification of viral HA and NP.

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Derivative Assay