Analysis Biomolecular Interactions of Antibodies to Influenza Virus Hemagglutinins by BLI (CAT#: STEM-MB-0117-CJ)

Name: Analysis Biomolecular Interactions of Antibodies to Influenza Virus Hemagglutinins by BLI
SKU: STEM-MB-0117-CJ
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Introduction

Influenza viruses are characterized by their rapid antigenic change as a result of their high mutation frequency. Therefore, the composition of influenza virus vaccines requires frequent updates (every 2 years on average for H3N2) to match their antigenicity as closely as possible to that of the variant viruses most prevalent in the population. Analysis of antibody responses that correlate with protective immunity to influenza virus vaccination is an important element in the assessment of the potential impact of viral antigenic drift. The hemagglutination inhibition (HI) test is the most widely used serological test for the detection of anti-influenza virus antibodies and is used routinely to determine the serological outcome of vaccinations.

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Principle Applications Procedure Materials

Principle

Bio-Layer Interferometry (BLI) is an optical technique for measuring macromolecular interactions by analyzing interference patterns of white light reflected from the surface of a biosensor tip. BLI experiments are used to determine the kinetics and affinity of molecular interactions. In a BLI experiment, one molecule is immobilized to a Dip and Read Biosensor and binding to a second molecule is measured. A change in the number of molecules bound to the end of the biosensor tip causes a shift in the interference pattern that is measured in real-time.

Applications

Immunology/Inflammation; Virology

Procedure

1. Detect Buffers and prepare samples. BLI experiments are set up with one molecule immobilised on the surface of the biosensor (load sample) and a second molecule in solution (the analytical sample).
2. Fix the load sample on the biocompatible biosensor while the analytical sample is in solution.
3. The biosensor tip is immersed in the solution so that the target molecule begins to bind to the analysis sample.
4. Set up and run the BLI experiment. Molecules bound to or dissociated from the biosensor can generate response curves on the BLI system; unbound molecules, changes in the refractive index of the surrounding medium or changes in flow rate do not affect the interferogram pattern.
5. Collect and analyse data on the BLI's system.