Analysis Biomolecular Interactions of Anti-Ebola mAb and Purified Glycoprotein (GP) by BLI (CAT#: STEM-MB-0133-CJ)

Introduction

The frequency and high mortality of Ebola virus outbreaks highlight the pressing need for diagnostic technologies, especially those that can be used for population surveillance. During public health emergencies, such as those induced by the four outbreaks of Ebola virus in the Democratic Republic of the Congo (DRC) that have occurred in the past 4 years. Antibody serology can also be used to survey a population to provide a more complete estimate of total infections, to begin to understand the immune response of survivors if it is not yet known, and to assess the success of virus containment to guide further control measures. Ebola virus antibodies can be detected by using the Ebola virus envelope glycoprotein ectodomain (GP), which lacks both the transmembrane and the mucin-like domain, as the affinity reagent.

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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

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: Chimeric Anti-EBOV GP mAb c6D8, Chimeric Anti-EBOV GP mAb h13F6, Mouse Anti-EBOV GP mAb 4F3.

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