Hemagglutinin Purification
Hemagglutinin Purification - Related Videos
12:18
Expression of Functional Recombinant Hemagglutinin and Neuraminidase Proteins from the Novel H7N9 Influenza Virus Using the Baculovirus Expression System
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Cited by 182 •
2013
Here we describe a way to express correctly folded and functional influenza virus surface antigens derived from the novel Chinese H7N9 virus in insect cells. The technique can be adapted to express ectodomains of any viral or cellular surface proteins.
10:32
A Miniaturized Glycan Microarray Assay for Assessing Avidity and Specificity of Influenza A Virus Hemagglutinins
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Cited by 9 •
2016
Using a printed glycan microarray strategy, a conventional 96-well plate assay was miniaturized for analysis of influenza A virus hemagglutinin avidity and specificity for sialic acid containing receptors.
04:47
A Method to Assess Fc-mediated Effector Functions Induced by Influenza Hemagglutinin Specific Antibodies
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Cited by 6 •
2018
We describe a method to measure the activation of Fc-mediated effector functions by antibodies that target the influenza virus hemagglutinin. This assay can also be adapted to assess the ability of monoclonal antibodies or polyclonal sera targeting other viral surface glycoproteins to induce Fc-mediated immunity.
08:19
Subnanometer-Resolution Structural Determination of Hemagglutinin from Cryo-Electron Tomography of Influenza Viruses
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Cited by 1 •
2025
This article presents a protocol for data processing of influenza viruses imaged using cryo-electron tomography and subsequent subtomogram averaging of the hemagglutinin glycoprotein. This protocol covers step-by-step data processing, from image preprocessing to final model refinement.
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
FAQs
These conditions must be controlled because hemagglutinin purification aims to retain the glycoprotein’s native structure. Changes in pH or temperature, and inappropriate detergent conditions, can affect structural preservation during separation. Maintaining suitable conditions helps produce material that remains informative for antibody characterization, viral binding studies, and other assays that depend on biologically relevant antigen properties.
The workflow can begin with viral particles, infected-cell lysates, or recombinant expression systems, and each source presents hemagglutinin in a different mixture of accompanying components. Clarification first reduces unwanted material, while chromatography provides a separation step for isolating the target glycoprotein. Thus, the source determines what must be separated before downstream analysis.
Native structural preservation helps maintain the features recognized by antibodies and involved in receptor binding. If those features are not retained, purified material may provide a less representative view of immune recognition or viral attachment. Structurally appropriate antigen therefore supports more meaningful studies of how antibodies recognize hemagglutinin and potentially block viral entry.
Purified hemagglutinin provides a defined antigen for examining differences in antibody recognition. Comparing responses to hemagglutinin preparations can help investigators characterize antigenic variation, meaning changes in antigenic features that influence immune detection. This defined material separates analysis of the viral surface glycoprotein from the complexity of whole viral particles or infected-cell mixtures.
A typical workflow starts by obtaining hemagglutinin from viral particles, infected-cell lysates, or a recombinant expression system. Clarification removes unwanted material from the starting preparation, followed by chromatography to separate hemagglutinin from remaining components. Throughout these stages, pH, temperature, and detergent conditions are controlled to support recovery of structurally preserved protein.
Monitoring pH, temperature, and detergent conditions is central to the process because these variables influence whether hemagglutinin retains its native structure during isolation. The purification workflow should maintain carefully controlled settings rather than treating separation as sufficient by itself. This attention improves the suitability of the resulting preparation for binding, antibody, and antigen-focused studies.
Purified hemagglutinin is useful when an experiment requires a defined antigen rather than the full complexity of viral particles or infected-cell material. It supports immunological assays, antibody characterization, and viral binding studies, allowing investigators to focus measurements on this surface glycoprotein. The preparation also enables analysis of antigenic variation in a controlled experimental context.
The process supplies a defined hemagglutinin antigen for vaccine development and quality assessment. In infection research, this material helps examine whether immune responses recognize features associated with viral attachment and whether antibodies can interfere with viral entry. Its defined composition also supports more consistent evaluation of antigen properties than analyses based only on complex starting mixtures.