Ha protein

Manufactured by Sino Biological

Sourced in China, United States

HA protein is a laboratory reagent used in various research applications. It is a glycoprotein that plays a crucial role in the structure and function of influenza viruses. The HA protein is responsible for the attachment and entry of the virus into host cells, making it an important tool for studying virus-host interactions and developing vaccines and therapeutics.

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Lab products found in correlation

Nunc immuno 96 well plates, by Thermo Fisher Scientific (4 mentions) Carbonate bicarbonate buffer, by Thermo Fisher Scientific (4 mentions) Ha specific monoclonal antibody, by Sino Biological (4 mentions) Accuskan fc microplate, by Thermo Fisher Scientific (4 mentions) Anti mouse igg hrp, by Thermo Fisher Scientific (4 mentions) Crystal violet, by Tianjin Damao (3 mentions) Endofit ovalbumin ova protein, by InvivoGen (2 mentions) A puerto rico 8 34, by Sino Biological (1 mentions) Agarose, by Thermo Fisher Scientific (1 mentions) Luciferase assay kit, by Promega (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)

7 protocols using ha protein

Immunization Protocol with Antigen and Chitosan Adjuvant

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All immunizations were performed under anesthesia using an isoflurane vaporizer. Immunizations were administered intramuscularly (i.m.) in 50 µl total volume. All immunizations contained either 5 µg EndoFit ovalbumin (OVA) protein (Invivogen) or 1 µg hemagglutinin (HA) recombinant protein from A/California/07/2009 H1N1 (pdm09) (International Reagent Resource), or 1 µg HA protein from A/Puerto Rico/8/34 (Sino Biological Inc.), as indicated. In addition to antigen, mice received LMW or HMW CS at 4 or 40 µg. Low dose, 1 µg antigen per mouse vaccinations was chosen to allow for modest CS MW effects to be observed, as well as to investigate potential antigen dose sparing effects of CS adjuvantation (Lampe et al., 2020). As a negative control, mice were immunized with antigen protein alone. Low dose PR8 immunizations were delivered intranasally (i.n.) at 500 egg infective dose (EID)₅₀ in 30 µl PBS as a positive control for protection against viral challenge and antibody production. Antigen combined with 20 µg MPLA delivered i.m. was also used as a positive control for antibody production (Lampe et al., 2020). Mice were weighed for up to 7 days after immunization to assess adverse effects caused by adjuvantation.

Lampe A.T., Farris E.J., Brown D.M, & Pannier A.K. (2020). High‐ and low‐molecular‐weight chitosan act as adjuvants during single‐dose influenza A virus protein vaccination through distinct mechanisms. Biotechnology and Bioengineering, 118(3), 1224-1243.

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MDCK Cell-based Influenza Assay

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Madin-Darby Canine Kidney (MDCK) cells were obtained from ATCC (Manassas, VA, USA), LinX-A cells were donated by the Scripps Research Institute in San Diego, CA, USA. Competent cells DH5a and Stb13 (Trans Gen BiotechCo., Ltd., Beijing, China). Eight days old SPF chicken embryos (DaHuaNong Biotech Co., Ltd., Guangzhou, China). DMEM and 0.25% Trypsin-EDTA (Gibco, Grand Island, NY, USA), Purelink Plasmid Extraction Kit (Invitrogen, Carlsbad, CA, USA), penicillin-streptomycin antibiotic (Gbico, USA), DMSO (SIGMA-ALDRICH, USA), Luciferase Assay Kit (Promega, Madison, WI, USA), Cellular Lysate for Luciferase Assay (Promega, USA), Lipofectamine^TM 3000 Transfection Reagent (Promega, USA), high fidelity enzyme and Dpn I enzyme (Takara, USA), Neuraminidase inhibitor screening reagent (Beyotime Biotech Co., Ltd., Shanghai, China), H5 antibody and H5 antigen (Harbin veterinary research institute, Harbin, China), HA protein (Sino Biological Inc., Beijing, China), Biotin Labeling Kit PN21343 (Thermo Scientific Pierce Protein Biology, USA), Agarose (Invitrogen, USA), Crystal Violet (Damao Chemical Reagent Factory, Tianjin, China).

Shi W.Z., Jiang L.Z., Song G.P., Wang S., Xiong P, & Ke C.W. (2020). Study on the Antiviral Activities and Hemagglutinin-Based Molecular Mechanism of Novel Chlorogenin 3-O-β-Chacotrioside Derivatives against H5N1 Subtype Viruses. Viruses, 12(3), 304.

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Quantifying Influenza Antibody Levels

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Nunc Immuno 96 well plates (Fisher Scientific) were coated with 1μg/ml (50 μl/well) HA protein (Sino Biological) diluted in carbonate/bicarbonate buffer (Fisher Scientific) overnight at 4°C or 1 hour at 37°C. After washing and blocking with TBS for 1 hour the serum samples were diluted and added to the plate. Serially diluted HA-specific monoclonal antibody (Sino Biological) served as standard. Anti-mouse IgG-HRP (1:20,000; Fisher Scientific) in combination with TMB (Fisher Scientific) solution was used for detection. The signals were read at 450 nm using accuSkan FC microplate photometer (Fisher Scientific).

Qin Z., Bouteau A., Herbst C, & Igyártó B.Z. (2022). Pre-exposure to mRNA-LNP inhibits adaptive immune responses and alters innate immune fitness in an inheritable fashion. PLoS Pathogens, 18(9), e1010830.

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ELISA for HA-specific Antibodies

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Nunc Immuno 96 well plates (Fisher Scientific) were coated with 1μg/ml (50 μl/well) HA protein (Sino Biological) diluted in carbonate/bicarbonate buffer (Fisher Scientific) overnight at 4 °C or 1 hour at 37 °C. After washing and blocking with TBS for 1 hour the serum samples were diluted and added to the plate. Serially diluted HA-specific monoclonal antibody (Sino Biological) served as standard. Anti-mouse IgG-HRP (1:20,000; Fisher Scientific) in combination with TMB (Fisher Scientific) solution was used for detection. The signals were read at 450 nm using accuSkan FC microplate photometer (Fisher Scientific).

Ndeupen S., Bouteau A., Herbst C., Qin Z., Jacobsen S., Powers N.E., Hutchins Z., Kurup D., Diba L.Z., Watson M., Ramage H, & Igyártó B.Z. (2022). Langerhans cells and cDC1s play redundant roles in mRNA-LNP induced protective anti-influenza and anti-SARS-CoV-2 immune responses. PLoS Pathogens, 18(1), e1010255.

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Quantitative HA Antibody ELISA

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Nunc Immuno 96 well plates (Fisher Scientific) were coated with 1μg/ml (50 μl/well) HA protein (Sino Biological) diluted in carbonate/bicarbonate buffer (Fisher Scientific) overnight at 4°C or 1 hour at 37 °C. After washing and blocking with TBS for 1 hour the serum samples were diluted and added to the plate. Serially diluted HA-specific monoclonal antibody (Sino Biological) served as standard. Anti-mouse IgG-HRP (1:20,000; Fisher Scientific) in combination with TMB (Fisher Scientific) solution was used for detection. The signals were read at 450 nm using accuSkan FC microplate photometer (Fisher Scientific).

Qin Z., Bouteau A., Herbst C, & Igyártó B.Z. (2022). Pre-exposure to mRNA-LNP inhibits adaptive immune responses and alters innate immune fitness in an inheritable fashion. bioRxiv.

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Adjuvant Effects of Chitosan on Influenza Vaccination

Cited 1 time

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All immunizations were performed under anesthesia using isoflurane vaporizer. Immunizations were administered intramuscularly (i.m.) in 50 μl total volume. All immunizations contained either 5 μg EndoFit Ovalbumin (OVA) protein (Invivogen) or 1 μg hemagglutinin (HA) recombinant protein from A/California/07/2009 H1N1 (pdm09) (International Reagent Resource, Manassas, VA, USA), or 1 μg HA protein from A/Puerto Rico/8/34 (Sino Biological Inc., Beijing, China), as indicated. In addition to antigen, mice received LMW or HMW CS at 4 or 40 μg. Low dose, 1 μg antigen per mouse vaccinations was chosen to allow for modest CS MW effects to be observed, as well as to investigate potential antigen dose sparing effects of CS adjuvantation (Lampe et al., 2020 (link)). As a negative control, mice were immunized with antigen protein alone. Low dose PR8 immunizations were delivered intranasally (i.n.) at 500 egg infective dose (EID)₅₀ in 30 μl PBS as a positive control for protection against viral challenge and antibody production. Antigen combined with 20 μg MPLA delivered i.m. was also used as a positive control for antibody production (Lampe et al., 2020 (link)). Mice were weighed for up to 7 days after immunization to assess adverse effects caused by adjuvantation.

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ELISA for HA-specific Antibodies

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Nunc Immuno 96 well plates (Fisher Scientific) were coated with 1μg/ml (50 μl/well) HA protein (Sino Biological) diluted in carbonate/bicarbonate buffer (Fisher Scientific) overnight at 4 °C or 1 hour at 37 °C. After washing and blocking with TBS for 1 hour the serum samples were diluted and added to the plate. Serially diluted HA-specific monoclonal antibody (Sino Biological) served as standard. Anti-mouse IgG-HRP (1:20,000; Fisher Scientific) in combination with TMB (Fisher Scientific) solution was used for detection. The signals were read at 450 nm using accuSkan FC microplate photometer (Fisher Scientific).

Ndeupen S., Bouteau A., Herbst C., Qin Z., Hutchins Z., Kurup D., Diba L.Z, & Igyártó B.Z. (2021). Langerhans cells and cDC1s play redundant roles in mRNA-LNP induced protective anti-influenza and anti-SARS-CoV-2 responses. bioRxiv.

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