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Analysis Biomolecular Interactions of the Hexahistidine-peptide with Chimeric VLPs by BLI (CAT#: STEM-MB-0136-CJ)
Introduction
Virus-like-particles (VLPs) are self-assembled, proteinaceous nanoparticles derived from naturally occurring viruses. In last years, VLPs attracted broad scientific interest due to their properties as versatile scaffold in nanotechnology. These beneficial characteristics are mainly originated from their particle integrity. Since VLPs are not infectious and do not replicate, they are considered as a safe format in biomedicine. Virus-like-particles per se are robust structures, and are highly immunogenic as the extremely repetitive and dense presentation of epitopes on the VLP surface efficiently stimulates B cells. The majorities of these alterations implicate disadvantages in vaccine development as they directly affect the foreign epitope sequence and may negatively change the epitope conformation.
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.
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: Human Aβ Peptides
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: Human Aβ Peptides
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