Analysis Biomolecular Interactions of DS003 to gp120 by BLI (CAT#: STEM-MB-0115-CJ)

Introduction

A major target for potential HIV pre-exposure prophylaxis (PrEP) intervention is the binding of viral gp120 to CD4+. Inhibiting this step can be achieved by blocking the CD4+-binding site within gp120. Small proteins mimicking CD4+ have shown high potency against HIV-1 in-vitro and in animal studies. An alternative to inhibit effective gp120-CD4+ binding is for a drug to bind to gp120 and block the conformational change in gp120 induced by CD4+. DS003 (BMS-599793) is another related compound with a similar mechanism of action, developed for topical PrEP as a microbicide candidate by the International Partnership for Microbicides (IPM). DS003 has been advanced to a Phase I clinical trial, IPM042, as a vaginal tablet showing a good safety profile and achieving concentrations in the female genital tract that were capable of inhibiting ex-vivo HIV-1 infection of cervical biopsies.

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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: Dulbecco's Minimal Essential Medium (DMEM)

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