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Analysis Biomolecular Interactions of A/H10 Seal Influenza Viruses with Avian-type Receptor Analogs by BLI (CAT#: STEM-MB-0164-CJ)

Introduction

Avian influenza A/H10N7 virus can infect caused an outbreak in harbor seals (Phoca vitulina) and gray seals (Halichoerus grypus) in North-European coastal waters. Phylogenetic analysis showed amino-acid sequence differences between the HA of seal A/H10N7 viruses and closely related Eurasian avian A/H10N7 viruses and the accumulation of amino-acid substitutions in seal A/H10N7 viruses over time. Since the known avian-origin A/H10N7 and A/H10N8 viruses have demonstrated their ability to infect humans, it is possible that the avian-origin mammalian-adapted seal A/H10N7 virus poses an even greater risk for humans.




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: NP monoclonal antibody (IgG2a, clone Hb65)