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Analysis Biomolecular Interactions of Antibody with Plasmodium Falciparum Circumsporozoite Protein (CSP) Antigens by BLI (CAT#: STEM-MB-0090-CJ)

Introduction

The Plasmodium falciparum circumsporozoite protein (CSP) is considered a major antimalarial vaccine target. CSP consists of an N-terminal domain, a central repeat region consisting of NANP and NVDP amino acid repeats, and a C-terminal region with a glycophosphatidylinositol anchor domain for plasma membrane anchorage. RTS,S, the most advanced malaria vaccine candidate, consists of 19 NANP repeats and the C-terminal region of CSP fused to the hepatitis B surface Ag. In humans, the immunogenicity assessments following RTS,S/AS01 immunization point toward a critical role for anti-CSP IgG Abs with specificity for the NANP repeat region and possibly CSP-specific CD4+ T cell responses.




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; Pharmacology; 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: Gator® Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: Virus Strains