Analysis Biomolecular Interactions of Broadly Neutralizing Antibodies (bNAbs) with 16055 SOSIP.v5.2 and 16055 SOSIP.v8.3 by BLI (CAT#: STEM-MB-0229-CJ)

Introduction

An effective vaccine requires the induction of broadly neutralising antibodies (bNAb). Generally, bNAbs emerge after years of virus-antibody co-evolution in infected individuals; they usually recognise conserved epitopes on densely glycosylated envelope glycoproteins (Env). The development of recombinant immunogens that closely resemble the natural trimeric conformation of Env, such as those designed on the basis of SOSIP, could accelerate the development of HIV-1 vaccines by triggering autologous NAb reactions in various animal models.Liposomes based on the natural flexible linkage (NFL) Env trimer of 16055; an evolved branching C virus isolate with a Tier-2 phenotype. These immunogens induce an autologous NAb response in rhesus monkeys and 16055 may be a suitable Env genotype.

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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

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: Fortebio Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: Modified 16055 SOSIP.v5.2 sequence

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