Analysis Biomolecular Interactions of H5 HA Specific MAbs with Fulllength H5N1 by BLI (CAT#: STEM-MB-0135-CJ)

Introduction

H5N1 viruses particularly cause high mortality in both birds and humans. Over 600 human cases with highly pathogenic avian influenza (HPAI) H5N1 viruses have been reported by the World Health Organization from several countries across the globe since November 2003 with approximately 60% fatality. H5N1 viruses have emerged by continual antigenic drift in poultry and wild bird populations with distinct geographical distribution. Airborne transmission of H5N1 virus in humans has not been previously reported and occurs upon long-term exposure of care givers to infected patients. A few mutations in H5N1 viruses, primarily in the receptor binding site (RBS) of H5 hemagglutinin (HA), enabled the virus to transmit via droplets in ferrets.

Monoclonal antibodies (MAbs) specifically generated against HPAI H5N1 viruses have the potential to be used in detection. Furthermore, neutralizing MAbs can prospectively be applied as an alternate strategy to combat influenza A (H5N1) virus, as prop

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Principle Applications Procedure Materials Notes Related Products

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; Pharmacology

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.

Materials

• Equipment: Fortebio Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: Fulllength H5N1 A/turkey/Turkey/1/2005 rHA

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