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Analysis Biomolecular Interactions of Human Papillomavirus Virus-Like Particles (VLPs) with Yeast Whole Cell RNA by BLI (CAT#: STEM-MB-0245-CJ)

Introduction

Virus-like particles (VLPs) are virus-resembling nanoparticles formed by viral structural proteins but do not have any core genetic material. Because VLPs are not infective, they are considered as one powerful tool for a broad spectrum of applications, including vaccines, antibody development, lipoparticle technology, delivery systems, bioimaging and cell targeting, etc.<br /><br />Human papillomavirus (HPV) is a small DNA virus from the Papillomaviridae family with a genome of about 8,000 base pairs. The HPV genome is composed of six early ORFs (E1, E2, E4, E5, E6, and E7), two late ORFs (L1 and L2), and a non-coding long control region (LCR). HPV can invades into the host cells by integrating genomes with the host genome. The genome integration disrupts E2 ORF thus preventing E2 repression on E6 and E7. Infection on host cells increases E6 and E7 expression which promotes cellular proliferation and the chance of malignancy.




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: ForteBio Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: Recombinant Human Papillomavirus (HPV) Type 11 and Type 18 VLPs