Analysis Biomolecular Interactions of mAbs witj HCV J6 E2 Glycoprotein by BLI (CAT#: STEM-MB-0179-CJ)

Introduction

HCV is a blood-borne virus and people who inject drugs have the highest risk for infection.Early appearance of serum nAbs is associated with spontaneous clearance of acute HCV infection and reinfection in humans. Elucidation of the crystal structure of HCV envelope glycoprotein E2, peptide mapping, and computerized modelling of antibodies against E1E2 heterodimer has led to identification of conserved nAb binding epitopes on E1 and E2. HCV E2 tetrimer decoys can be used to study phenotypes, gene expression patterns, B-cell receptor (BCR) libraries, and function of antigen-specific MBCs in patients who spontaneously resolve acute HCV infection (defectors) or chronic infection after drug injection.

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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: Gator® Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: Directed Mutagenesis Kit

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