Analysis Kinetics of SARS-CoV-2 Truncated Aptamer by BLI (CAT#: STEM-MB-0284-CJ)

Introduction

Aptamers are synthetic nucleic acid sequences selected for their affinity to specifically bind molecular targets of choice, including small molecules, proteins, and pathogens. They are selected from a large library of randomly generated sequences through iterations of increasing selective pressure. They have numerous advantages over other affinity binding agents such as antibodies, including chemical stability (improved shelf life), the ability to synthesize large quantities at a relatively low cost, and the easy functionalization with chemical groups for labeling, surface immobilization, or detection. Consequently, they are very compatible with electrochemical measurements.

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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; 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: Thin-film gold electrodes (TFGEs), Molecular biology water, Cytiva illustra NAP Columns (NAP-10), Potassium Ferricyanide, Potassium Ferrocyanide, Potassium Chloride, Sodium Sulfate

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