recombinant influenza ha proteins  (Sino Biological)


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    Structured Review

    Sino Biological recombinant influenza ha proteins
    Increased strain-specific IgG titers after a single MHCII-targeted vaccination. BALB/c mice ( n = 6 per group) were immunized once with 25 μg of plasmids encoding one of the indicated vaccines. ( A ) Sera were harvested at weeks 2, 3, and 4 after vaccination, and IgG responses in sera from each of the vaccine groups were measured by ELISAs against <t>recombinant</t> <t>HA</t> from H5, H6, H8, H9, H11, H13, and H1 (PR8) <t>influenza</t> viruses (mean ± SEM). ** p
    Recombinant Influenza Ha Proteins, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant influenza ha proteins/product/Sino Biological
    Average 94 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    recombinant influenza ha proteins - by Bioz Stars, 2022-10
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    Images

    1) Product Images from "Simultaneous Targeting of Multiple Hemagglutinins to APCs for Induction of Broad Immunity against Influenza"

    Article Title: Simultaneous Targeting of Multiple Hemagglutinins to APCs for Induction of Broad Immunity against Influenza

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1701088

    Increased strain-specific IgG titers after a single MHCII-targeted vaccination. BALB/c mice ( n = 6 per group) were immunized once with 25 μg of plasmids encoding one of the indicated vaccines. ( A ) Sera were harvested at weeks 2, 3, and 4 after vaccination, and IgG responses in sera from each of the vaccine groups were measured by ELISAs against recombinant HA from H5, H6, H8, H9, H11, H13, and H1 (PR8) influenza viruses (mean ± SEM). ** p
    Figure Legend Snippet: Increased strain-specific IgG titers after a single MHCII-targeted vaccination. BALB/c mice ( n = 6 per group) were immunized once with 25 μg of plasmids encoding one of the indicated vaccines. ( A ) Sera were harvested at weeks 2, 3, and 4 after vaccination, and IgG responses in sera from each of the vaccine groups were measured by ELISAs against recombinant HA from H5, H6, H8, H9, H11, H13, and H1 (PR8) influenza viruses (mean ± SEM). ** p

    Techniques Used: Mouse Assay, Recombinant

    Ab responses against all included HAs after DNA immunization with vaccine mixtures. BALB/c mice ( n = 6 per group) were immunized three times (weeks 0, 4, and 12) with 25 μg of DNA of the indicated vaccine (arrowheads). Serum samples were harvested 2–3 wk after each vaccine delivery, and HA-specific IgG responses were measured in sandwich ELISAs against recombinant HA from H5, H6, H8, H9, H11, and H13 influenza viruses (mean ± SEM). * p
    Figure Legend Snippet: Ab responses against all included HAs after DNA immunization with vaccine mixtures. BALB/c mice ( n = 6 per group) were immunized three times (weeks 0, 4, and 12) with 25 μg of DNA of the indicated vaccine (arrowheads). Serum samples were harvested 2–3 wk after each vaccine delivery, and HA-specific IgG responses were measured in sandwich ELISAs against recombinant HA from H5, H6, H8, H9, H11, and H13 influenza viruses (mean ± SEM). * p

    Techniques Used: Mouse Assay, Recombinant

    2) Product Images from "Pan-Influenza A Protection by Prime–Boost Vaccination with Cold-Adapted Live-Attenuated Influenza Vaccine in a Mouse Model"

    Article Title: Pan-Influenza A Protection by Prime–Boost Vaccination with Cold-Adapted Live-Attenuated Influenza Vaccine in a Mouse Model

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.00116

    Cross-reactive hemagglutinin (HA)-specific antibody responses elicited by vaccination. (A) Breadth of the HA-specific sera IgG antibodies. To examine the breadth of HA-specific sera IgG antibodies induced by vaccination, recombinant HA proteins expressed in insect cells were used as coating antigens in ELISA. The HA proteins tested include five different group 1 HAs from H1N1 (A/California/6/2009), H1N1 (A/Puerto Rico/8/1934), H2N2 (A/Canada/720/2006), H5N1 (A/Indonesia/5/2005), and H9N2 (A/Hong Kong/35820/2009) and two group 2 HAs from H3N2 (A/Sydney/5/1997) and H7N9 (A/Anhui/1/2013) influenza viruses. (B) Antibody titers specific to HA full-length or stalk of pH1N1 or PR8 (H1N1) virus. Using the Escherichia coli -expressed LysRS-HA fusion proteins as coating antigens, sera IgG antibodies specific to the HA full-length or stalk protein were measured by ELISA. Antibody titers were expressed as the reciprocal serum dilution that yielded OD 450 greater than the mean + 2 SD (SD) of PBS control group. Data are the mean of each cohort ( N = 5), and error bars indicate SD. *** P
    Figure Legend Snippet: Cross-reactive hemagglutinin (HA)-specific antibody responses elicited by vaccination. (A) Breadth of the HA-specific sera IgG antibodies. To examine the breadth of HA-specific sera IgG antibodies induced by vaccination, recombinant HA proteins expressed in insect cells were used as coating antigens in ELISA. The HA proteins tested include five different group 1 HAs from H1N1 (A/California/6/2009), H1N1 (A/Puerto Rico/8/1934), H2N2 (A/Canada/720/2006), H5N1 (A/Indonesia/5/2005), and H9N2 (A/Hong Kong/35820/2009) and two group 2 HAs from H3N2 (A/Sydney/5/1997) and H7N9 (A/Anhui/1/2013) influenza viruses. (B) Antibody titers specific to HA full-length or stalk of pH1N1 or PR8 (H1N1) virus. Using the Escherichia coli -expressed LysRS-HA fusion proteins as coating antigens, sera IgG antibodies specific to the HA full-length or stalk protein were measured by ELISA. Antibody titers were expressed as the reciprocal serum dilution that yielded OD 450 greater than the mean + 2 SD (SD) of PBS control group. Data are the mean of each cohort ( N = 5), and error bars indicate SD. *** P

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay

    Experimental design of vaccination and challenge in mice. (A) Vaccination schedule and design of prime–boost vaccination against heterologous challenge. Four different prime–boost vaccination groups were designed using three different cold-adapted, live attenuated influenza vaccines (CAIVs), ca-pH1N1, ca-NCH1N1, and ca-IDH5N1. Prime and boost CAIVs (10 5 PFUs of each vaccine) were administered into mice via intranasal route with two weeks interval. A month later, each group was divided into four subgroups (20 subgroups in total) and challenged with 10 mouse lethal dose 50 (MLD 50 ) of each of four heterologous influenza viruses. (B) Phylogenetic tree of influenza A hemagglutinin (HA) proteins. HA to which binding affinity of vaccination-induced antibodies or protection efficacy in vivo tested in this study was highlighted in colored circles.
    Figure Legend Snippet: Experimental design of vaccination and challenge in mice. (A) Vaccination schedule and design of prime–boost vaccination against heterologous challenge. Four different prime–boost vaccination groups were designed using three different cold-adapted, live attenuated influenza vaccines (CAIVs), ca-pH1N1, ca-NCH1N1, and ca-IDH5N1. Prime and boost CAIVs (10 5 PFUs of each vaccine) were administered into mice via intranasal route with two weeks interval. A month later, each group was divided into four subgroups (20 subgroups in total) and challenged with 10 mouse lethal dose 50 (MLD 50 ) of each of four heterologous influenza viruses. (B) Phylogenetic tree of influenza A hemagglutinin (HA) proteins. HA to which binding affinity of vaccination-induced antibodies or protection efficacy in vivo tested in this study was highlighted in colored circles.

    Techniques Used: Mouse Assay, Binding Assay, In Vivo

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    Sino Biological recombinant mex4108 ha protein plasma
    Validated miRNA targets are involved in immune response and cell proliferation in the lungs. Expression levels of (A) KRAS , BLIMP1 , and CDK6 (targets of let-7f); (B) SIRT1 , ZAP70 , and MYC (targets of miR-34c); (C) CAMTA1 (target of miR-129); and (D) PTEN (target of miR-18b) in BAL from rhesus macaques (five aged and three young adults) following <t>MEX4108</t> infection are shown. Expression of mRNA was normalized to expression of RPL32 . Changes in gene expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. (A–D) *let-7f, miR-34c, miR-129, miR-18-b, KRAS , SIRT1 , CAMTA1 , and PTEN ; ‡ KRAS and ZAP70 ; † CDK6 and MYC . * p
    Recombinant Mex4108 Ha Protein Plasma, supplied by Sino Biological, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant mex4108 ha protein plasma/product/Sino Biological
    Average 86 stars, based on 1 article reviews
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    94
    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
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    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    94
    Sino Biological h3n2 a brisbane 10 2007
    ELISA on Chip antigen dilution titer curves. (A) Monoclonal antibody (mAb) titer curves: Example titer curves of the binding level of 4 concentrations of the <t>anti-H3N2</t> Brisbane 2007 mAb with 11 serial 2-fold dilutions of <t>the</t> <t>spotted</t> <t>A/Brisbane/10/2007</t> rHA antigen. Each dilution of the mAb was incubated with a different antigen microarray (AM). (B) Three-dimensional titer curves: mAb titer curves for antibody serial dilution and spotted antigen serial dilution.
    H3n2 A Brisbane 10 2007, 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/h3n2 a brisbane 10 2007/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    h3n2 a brisbane 10 2007 - by Bioz Stars, 2022-10
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    Image Search Results


    Validated miRNA targets are involved in immune response and cell proliferation in the lungs. Expression levels of (A) KRAS , BLIMP1 , and CDK6 (targets of let-7f); (B) SIRT1 , ZAP70 , and MYC (targets of miR-34c); (C) CAMTA1 (target of miR-129); and (D) PTEN (target of miR-18b) in BAL from rhesus macaques (five aged and three young adults) following MEX4108 infection are shown. Expression of mRNA was normalized to expression of RPL32 . Changes in gene expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. (A–D) *let-7f, miR-34c, miR-129, miR-18-b, KRAS , SIRT1 , CAMTA1 , and PTEN ; ‡ KRAS and ZAP70 ; † CDK6 and MYC . * p

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: Validated miRNA targets are involved in immune response and cell proliferation in the lungs. Expression levels of (A) KRAS , BLIMP1 , and CDK6 (targets of let-7f); (B) SIRT1 , ZAP70 , and MYC (targets of miR-34c); (C) CAMTA1 (target of miR-129); and (D) PTEN (target of miR-18b) in BAL from rhesus macaques (five aged and three young adults) following MEX4108 infection are shown. Expression of mRNA was normalized to expression of RPL32 . Changes in gene expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. (A–D) *let-7f, miR-34c, miR-129, miR-18-b, KRAS , SIRT1 , CAMTA1 , and PTEN ; ‡ KRAS and ZAP70 ; † CDK6 and MYC . * p

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Expressing, Infection

    Differentially expressed miRNAs in PBMCs following MEX4108. Expression levels of miR-18b (A) , miR-20a (B) , miR-192 (C) , miR-451 (D) , miR-138 (E) , miR-193b (F) , miR-132 (G) , let-7f (H) , and miR-146b (I) in PBMCs from rhesus macaques ( n = 11; five aged and six young adults) following MEX4108 infection were determined using qRT-PCR. Expression of miRNAs was normalized to expression of U6 snRNA. Changes in microRNA expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. * p

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: Differentially expressed miRNAs in PBMCs following MEX4108. Expression levels of miR-18b (A) , miR-20a (B) , miR-192 (C) , miR-451 (D) , miR-138 (E) , miR-193b (F) , miR-132 (G) , let-7f (H) , and miR-146b (I) in PBMCs from rhesus macaques ( n = 11; five aged and six young adults) following MEX4108 infection were determined using qRT-PCR. Expression of miRNAs was normalized to expression of U6 snRNA. Changes in microRNA expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. * p

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Expressing, Infection, Quantitative RT-PCR

    MEX4108 infection results in increased frequency of pDCs and robust production of cytokine, chemokine, and growth factor levels in the lungs. (A) The frequencies of DCs (lin − CD14 − HLA-DR + ) and macrophages/monocytes (lin − CD14 + HLA-DR − ) in BAL cells were measured by flow cytometry. (B) The frequencies of mDC (CD123 − CD11c + ) and pDC (CD123 + CD11c − ) in BAL cells were measured by flow cytometry. Longitudinal analyses of the frequency of immune cells within BAL were carried out using a one-way repeated-measures ANOVA model, followed by Dunnett's multiple comparison post-test to explore differences between days postinfection and baseline (day 0) values. Mean ± SEM are shown. * p

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: MEX4108 infection results in increased frequency of pDCs and robust production of cytokine, chemokine, and growth factor levels in the lungs. (A) The frequencies of DCs (lin − CD14 − HLA-DR + ) and macrophages/monocytes (lin − CD14 + HLA-DR − ) in BAL cells were measured by flow cytometry. (B) The frequencies of mDC (CD123 − CD11c + ) and pDC (CD123 + CD11c − ) in BAL cells were measured by flow cytometry. Longitudinal analyses of the frequency of immune cells within BAL were carried out using a one-way repeated-measures ANOVA model, followed by Dunnett's multiple comparison post-test to explore differences between days postinfection and baseline (day 0) values. Mean ± SEM are shown. * p

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Infection, Flow Cytometry, Cytometry

    Model of differentially expressed microRNAs, gene targets, and immune response in the lungs following MEX4108 infection.

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: Model of differentially expressed microRNAs, gene targets, and immune response in the lungs following MEX4108 infection.

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Infection

    Pandemic H1N1 virus replicates to similar levels in young and aged macaques. Viral loads were measured using qRT-PCR using primers and probes specific for MEX4108 hemagglutinin (HA) in throat swabs (A) , nasal swabs (B) , ocular swabs (C) , and BAL fluid (D) . Viral genome copy number data were log transformed with base 10 and longitudinal changes of viral genome copy number between aged and young adults were compared using a two-way ANOVA, followed by Bonferroni's multiple comparison post-test to determine differences in viral load. Longitudinal changes were compared using repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test to explore differences between days postinfection and baseline (day 0) values, mean ± SEM are shown. (A–D) * for aged animals; ‡ for young adult animals; *** ,‡‡ p

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: Pandemic H1N1 virus replicates to similar levels in young and aged macaques. Viral loads were measured using qRT-PCR using primers and probes specific for MEX4108 hemagglutinin (HA) in throat swabs (A) , nasal swabs (B) , ocular swabs (C) , and BAL fluid (D) . Viral genome copy number data were log transformed with base 10 and longitudinal changes of viral genome copy number between aged and young adults were compared using a two-way ANOVA, followed by Bonferroni's multiple comparison post-test to determine differences in viral load. Longitudinal changes were compared using repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test to explore differences between days postinfection and baseline (day 0) values, mean ± SEM are shown. (A–D) * for aged animals; ‡ for young adult animals; *** ,‡‡ p

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Quantitative RT-PCR, Transformation Assay

    Differentially expressed miRNAs in BAL cells following MEX4108. Expression levels of let-7f (A) , miR-34c (B) , miR-129 (C) , miR-18b (D) , miR-146b (E) , miR-132 (F) , miR-192 (G) , and miR-138 (H) in BAL cells from rhesus macaques ( n = 8; five aged and three young adults) following MEX4108 infection were determined using qRT-PCR. Expression of miRNAs was normalized to expression of U6 snRNA. Changes in microRNA expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. * p

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: Differentially expressed miRNAs in BAL cells following MEX4108. Expression levels of let-7f (A) , miR-34c (B) , miR-129 (C) , miR-18b (D) , miR-146b (E) , miR-132 (F) , miR-192 (G) , and miR-138 (H) in BAL cells from rhesus macaques ( n = 8; five aged and three young adults) following MEX4108 infection were determined using qRT-PCR. Expression of miRNAs was normalized to expression of U6 snRNA. Changes in microRNA expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. * p

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Expressing, Infection, Quantitative RT-PCR

    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

    ELISA on Chip antigen dilution titer curves. (A) Monoclonal antibody (mAb) titer curves: Example titer curves of the binding level of 4 concentrations of the anti-H3N2 Brisbane 2007 mAb with 11 serial 2-fold dilutions of the spotted A/Brisbane/10/2007 rHA antigen. Each dilution of the mAb was incubated with a different antigen microarray (AM). (B) Three-dimensional titer curves: mAb titer curves for antibody serial dilution and spotted antigen serial dilution.

    Journal: medRxiv

    Article Title: ELISA–on-Chip: High throughput antibody profiling using antigen microarrays

    doi: 10.1101/2022.07.05.22277251

    Figure Lengend Snippet: ELISA on Chip antigen dilution titer curves. (A) Monoclonal antibody (mAb) titer curves: Example titer curves of the binding level of 4 concentrations of the anti-H3N2 Brisbane 2007 mAb with 11 serial 2-fold dilutions of the spotted A/Brisbane/10/2007 rHA antigen. Each dilution of the mAb was incubated with a different antigen microarray (AM). (B) Three-dimensional titer curves: mAb titer curves for antibody serial dilution and spotted antigen serial dilution.

    Article Snippet: These included three seasonal vaccine strains (north hemisphere): H3N2 A/Wisconsin/67/2005; H3N2 A/Brisbane/10/2007 and H1N1 A/California/07/2009, and the avian influenza H7N9 A/Shanghai/1/2013 strain (EoC AMs in ).

    Techniques: Enzyme-linked Immunosorbent Assay, Chromatin Immunoprecipitation, Binding Assay, Incubation, Microarray, Serial Dilution

    Magnitude, breadth and specificity of anti-influenza HA IgG response to A/PuertoRico/8/1934 (PR8) sublethal infection in mice, as measured by antigen microarrays spotted with recombinant HA (rH) proteins from 41 influenza strains. Nine C57BL/6 mice were infected intranasally (i.n.) with A/PuertoRico/8/1934 (PR8) H1N1 viruses, as previously described [ 13 ]. Serum samples were collected pre-infection (blue) and 28 days post infection (orange). Antigen microarrays spotted with 41 recombinant full HA proteins or HA1 units of 36 influenza A strains from 3 subtypes (11 rH1 proteins from H1N1 strains, including the PR8 infection strain; 18 rH3 proteins from H3N2 strains; and 7 rH5 proteins from H5N1 strains), and 5 influenza B strains, were used to measure IgG binding to each spotted protein by mean fluorescence intensity (MFI). (A) IgG binding to the rHA protein of the infection strain PR8 (MFI). (B) Geometric mean magnitudes (GMT) of IgG MFI of all the spotted proteins from each subtype. In panels A and B horizontal lines represent the median, boxes denote the 25th and 75th percentiles, and the error bars represent 1.5 times the interquartile range. Statistical significance was assessed using the Wilcoxon signed rank test (pre vs. post): * p

    Journal: medRxiv

    Article Title: ELISA–on-Chip: High throughput antibody profiling using antigen microarrays

    doi: 10.1101/2022.07.05.22277251

    Figure Lengend Snippet: Magnitude, breadth and specificity of anti-influenza HA IgG response to A/PuertoRico/8/1934 (PR8) sublethal infection in mice, as measured by antigen microarrays spotted with recombinant HA (rH) proteins from 41 influenza strains. Nine C57BL/6 mice were infected intranasally (i.n.) with A/PuertoRico/8/1934 (PR8) H1N1 viruses, as previously described [ 13 ]. Serum samples were collected pre-infection (blue) and 28 days post infection (orange). Antigen microarrays spotted with 41 recombinant full HA proteins or HA1 units of 36 influenza A strains from 3 subtypes (11 rH1 proteins from H1N1 strains, including the PR8 infection strain; 18 rH3 proteins from H3N2 strains; and 7 rH5 proteins from H5N1 strains), and 5 influenza B strains, were used to measure IgG binding to each spotted protein by mean fluorescence intensity (MFI). (A) IgG binding to the rHA protein of the infection strain PR8 (MFI). (B) Geometric mean magnitudes (GMT) of IgG MFI of all the spotted proteins from each subtype. In panels A and B horizontal lines represent the median, boxes denote the 25th and 75th percentiles, and the error bars represent 1.5 times the interquartile range. Statistical significance was assessed using the Wilcoxon signed rank test (pre vs. post): * p

    Article Snippet: These included three seasonal vaccine strains (north hemisphere): H3N2 A/Wisconsin/67/2005; H3N2 A/Brisbane/10/2007 and H1N1 A/California/07/2009, and the avian influenza H7N9 A/Shanghai/1/2013 strain (EoC AMs in ).

    Techniques: Infection, Mouse Assay, Recombinant, Binding Assay, Fluorescence

    ELISA and ELISA-on-Chip titer curves Pearson correlation. (A) Monoclonal antibody titer curves: binding of 15 serial 2-fold dilutions of anti-A/Brisbane/10/2007 rHA monoclonal antibody to the A/Brisbane/10/2007 rHA protein at a single concentration (4 or 62.5 μg/ml), as measured by ELISA (blue) or ELISA on Chip (red), respectively. The curves were fitted using a 5 parameter logistic regression model. The Pearson correlation between the two curves was: r = 0.975, p = 7.1x10^ (-10). (B) Human samples titer curves correlation: IgG antibodies against 4 Influenza strains were quantified in 10 healthy adult individuals using both ELISA and ELISA-on-Chip assays. ELISA was performed over 15 serial (2-fold) sample dilutions using a single antigen concentration (4 μg/ml). The ELISA-on-Chip was run against antigens spotted in 11 serial dilutions (2-fold, as listed in the Y axis). Pearson correlation coefficients were computed for titer curves for each antigen concentration in ELISA-on- chip compared with the single antigen concentration in ELISA.

    Journal: medRxiv

    Article Title: ELISA–on-Chip: High throughput antibody profiling using antigen microarrays

    doi: 10.1101/2022.07.05.22277251

    Figure Lengend Snippet: ELISA and ELISA-on-Chip titer curves Pearson correlation. (A) Monoclonal antibody titer curves: binding of 15 serial 2-fold dilutions of anti-A/Brisbane/10/2007 rHA monoclonal antibody to the A/Brisbane/10/2007 rHA protein at a single concentration (4 or 62.5 μg/ml), as measured by ELISA (blue) or ELISA on Chip (red), respectively. The curves were fitted using a 5 parameter logistic regression model. The Pearson correlation between the two curves was: r = 0.975, p = 7.1x10^ (-10). (B) Human samples titer curves correlation: IgG antibodies against 4 Influenza strains were quantified in 10 healthy adult individuals using both ELISA and ELISA-on-Chip assays. ELISA was performed over 15 serial (2-fold) sample dilutions using a single antigen concentration (4 μg/ml). The ELISA-on-Chip was run against antigens spotted in 11 serial dilutions (2-fold, as listed in the Y axis). Pearson correlation coefficients were computed for titer curves for each antigen concentration in ELISA-on- chip compared with the single antigen concentration in ELISA.

    Article Snippet: These included three seasonal vaccine strains (north hemisphere): H3N2 A/Wisconsin/67/2005; H3N2 A/Brisbane/10/2007 and H1N1 A/California/07/2009, and the avian influenza H7N9 A/Shanghai/1/2013 strain (EoC AMs in ).

    Techniques: Enzyme-linked Immunosorbent Assay, Chromatin Immunoprecipitation, Binding Assay, Concentration Assay

    ELISA and AM correlations. We compared the area under the curve of 10 healthy adult individuals using our influenza antigen microarrays (diluted 1:3200, blue) and using traditional ELISA in a single antigen concentration (orange). The Spearman correlation between the AUC scores was computed for all pairwise combinations of four influenza strains: two H3N2 strains A/Wisconsin/67/2005, A/Brisbane/10/2007, the H1N1 A/California/07/2009 and the H7N9 A/Shanghai/1/2013

    Journal: medRxiv

    Article Title: ELISA–on-Chip: High throughput antibody profiling using antigen microarrays

    doi: 10.1101/2022.07.05.22277251

    Figure Lengend Snippet: ELISA and AM correlations. We compared the area under the curve of 10 healthy adult individuals using our influenza antigen microarrays (diluted 1:3200, blue) and using traditional ELISA in a single antigen concentration (orange). The Spearman correlation between the AUC scores was computed for all pairwise combinations of four influenza strains: two H3N2 strains A/Wisconsin/67/2005, A/Brisbane/10/2007, the H1N1 A/California/07/2009 and the H7N9 A/Shanghai/1/2013

    Article Snippet: These included three seasonal vaccine strains (north hemisphere): H3N2 A/Wisconsin/67/2005; H3N2 A/Brisbane/10/2007 and H1N1 A/California/07/2009, and the avian influenza H7N9 A/Shanghai/1/2013 strain (EoC AMs in ).

    Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay

    Subtype specificity of influenza rHA microarrays. Three groups of mice (n=9-10 per group) were injected intraperitoneally (IP) with sublethal doses of influenza A viruses from 3 different subtypes: pandemic A/California/07/2009 (Cal09, HlNl), A/X31/1968 (X31, H3N2) and A/Vietnam/1203/2004 (Viet1203, HSNl). Serum was collected at day 56 post infection and lgG antibodies binding to the recombinant hemagglutinin (rH) proteins were profiled using an influenza antigen microarray. (A) lgG responses to HlNl strains (rHl n=lO), (B) lgG responses to H3N2 strains (rH3, n=17), and (C) lgG responses to HSNl strains (rHS, n=6). Each bar represents the cumulative MFI for a single mouse to all the rH proteins listed. Response to the vaccine strain is in orange.

    Journal: medRxiv

    Article Title: ELISA–on-Chip: High throughput antibody profiling using antigen microarrays

    doi: 10.1101/2022.07.05.22277251

    Figure Lengend Snippet: Subtype specificity of influenza rHA microarrays. Three groups of mice (n=9-10 per group) were injected intraperitoneally (IP) with sublethal doses of influenza A viruses from 3 different subtypes: pandemic A/California/07/2009 (Cal09, HlNl), A/X31/1968 (X31, H3N2) and A/Vietnam/1203/2004 (Viet1203, HSNl). Serum was collected at day 56 post infection and lgG antibodies binding to the recombinant hemagglutinin (rH) proteins were profiled using an influenza antigen microarray. (A) lgG responses to HlNl strains (rHl n=lO), (B) lgG responses to H3N2 strains (rH3, n=17), and (C) lgG responses to HSNl strains (rHS, n=6). Each bar represents the cumulative MFI for a single mouse to all the rH proteins listed. Response to the vaccine strain is in orange.

    Article Snippet: These included three seasonal vaccine strains (north hemisphere): H3N2 A/Wisconsin/67/2005; H3N2 A/Brisbane/10/2007 and H1N1 A/California/07/2009, and the avian influenza H7N9 A/Shanghai/1/2013 strain (EoC AMs in ).

    Techniques: Mouse Assay, Injection, Infection, Binding Assay, Recombinant, Microarray