Analysis Kinetics of Anti-VEGF165 Aptamer Sequence by BLI (CAT#: STEM-MB-0266-CJ)

Introduction

Vascular endothelial growth factor (VEGF) is a strong endothelial-cell-specific mitogen that plays central roles in the regulation of physiologic and pathologic angiogenesis. It can bind to extracellular receptor tyrosine kinases and activate different intracellular signaling pathways, which will in turn stimulate vascular endothelial cell growth, survival, and proliferation. In mammals, the VEGF family consists of five subgroups: VEGF-A, VEGF-B, VEGF-C, VEGF-D, and the placenta growth factor. VEGF-A is the prototype member, with at least four isoforms: VEGF121, VEGF165, VEGF189, and VEGF206. Among these, VEGF165 is the most potent pro-angiogenic isoform and has been widely used as a biomarker for several human diseases, including cancer, rheumatoid arthritis, bronchial asthma, and diabetic eye disease.

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Principle Applications Procedure Materials Notes Related Products

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

Oncology & Cancer; Immunology/Inflammation; 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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