Analysis Kinetics of Aptamer SEQ9 (A), SEQ9M (B), SEQ9MT (C) to ANGPTL4 by BLI (CAT#: STEM-MB-0269-CJ)

Introduction

Angiopoietin-like 4 (ANGPTL4) is an emerging therapeutic and diagnostic target for DR. Cell-based, animal, and clinical studies have collectively identified ANGPTL4 as a specific biomarker for DR. Its concentration level in aqueous humor and serum has a significant dose-dependent relationship with disease severity. Accurate monitoring of ANGPTL4 provides an effective tool for DR screening, diagnosis, treatment, and management.Tremendous effort has been devoted to developing cost-efficient and convenient diagnostic techniques for detection of disease-specific protein biomarkers.

Aptamer has low immunogenicity and toxicity; they can be prepared by chemical synthesis and are easy to modify and label; they are less affected by environmental factors, such as pH and temperature; they have structural stability; and they have a wide range of recognition targets and strong
specificity.

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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

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
• Optionals: Recombinant human angiopoietin-like 4 (ANGPTL4, His Tag)

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