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Analysis Biomolecular Interactions of the AMC008 SOSIP Trimer-induced NAbs with Non-NAbs by BLI (CAT#: STEM-MB-0209-CJ)

Introduction

SOSIP trimers have generally elicited strong autologous NAb responses, but only sporadic, inconsistent, and weak cross-NAb responses.The studies characterizing MAbs and bulk serum of SOSIP Env trimer-immunized rabbits and macaques showed that the Ab responses frequently target strain-specific glycan holes. Many vaccine-induced NAbs target epitopes that overlap those of non-NAbs. SOSIP-induced NAbs have been discovered that target the same epitope as well-known bNAbs elicited after natural HIV-1 infection, but without displaying the same potency or breadth.




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; 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
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