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Analysis Biomolecular Interactions of Monkeypox Virus (MPXV) A29 Protein with Monoclonal Antibody by BLI (CAT#: STEM-MB-0123-CJ)

Introduction

Orthopoxviruses are a genus of DNA viruses of which four species are known to cause disease in humans: vaccinia virus (VACV), cowpox virus (CPXV), variola virus (VARV), and monkeypox virus (MPXV).<br /><br />Monkeypox virus is a zoonotic virus, endemic to central and Western Africa that can cause smallpox-like symptoms in humans.The virus was initially identified in non-human primate rash lesions in 1958 and first identified in humans in 1970 during the smallpox eradication efforts. Incidence of MPXV infections in the Democratic Republic of the Congo (DRC), where the majority of cases occur, has increased as much as 20-fold since the end of smallpox vaccination in 1980. The first case of human MPXV in the USA occurred in 2003 and was linked to the handling of prairie dogs that had been housed with imported African rodents.Similar to VARV or smallpox, MPXV disease presents with fever 10–12 days after exposure, followed by rash 2–3 days later. This rash progresses from macular to papular, vesicular, and finally pustular phases, similar to smallpox. This makes differentiation between these diseases based on clinical presentation difficult. While smallpox was declared eradicated by the World Health Organization in 1979, it is still a concern as a potential biological weapon or potential accidental release.




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

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