Analysis Biomolecular Interactions of Monoclonal Mouse Anti-PA Abs with PA by BLI (CAT#: STEM-MB-0147-CJ)

Introduction

Bacillus anthracis, the etiological agent of anthrax is one of the major bioterrorism-associated bacteria.Inhaled anthrax is the most hazardous form of exposure leading to bacteremia and to the secretion of bacterial exotoxins, i.e. protective antigen (PA) edema and lethal factors that induce tissue damage and even death in untreated cases. Therefore early diagnosis of anthrax is essential to prevent public catastrophe and allow an effective treatment. Diagnosis of anthrax can be achieved either by detection of the bacterial spore in contaminated fields or by monitoring the levels of the bacterial toxin—PA in clinical serum samples that was in high correlation with bacteremia.

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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, Toxicology

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: Mouse IgG Anti-PA Monoclonal Abs (Mab2, Mab3, Mab5)

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