Unlock Exclusive Discounts & Flash Sales! Click Here to Join the Deals on Every Wednesday!

Analysis Biomolecular Interactions of the Enzymatic Inhibitors to Factor D by BLI (CAT#: STEM-MB-0120-CJ)

Introduction

Several enzymes playing important roles in complement activation by processing the complement proteins. Among them, the plasma concentration of complement factor D (denoted as factor D) was reported to be the lowest among all complement related proteins and considered a rate-limiting enzyme. Acting on the upstream of AP, factor D cleaves complement factor B bound with C3b into the C3bBb complex11 . The ubiquitous role of factor D in AP activation led to the implication that inhibition of factor D can be an attractive strategy in the complement targeted therapy. Factor D belongs to the serine protease family12. Previous factor D inhibitor developments have led to the discovery of non-specific catalytic site inhibitors including coumarin derived covalent inhibitors.




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

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
Advertisement