Analysis Biomolecular Interactions of DNA Aptamers with Marine Biotoxin Gymnodimine-A by BLI (CAT#: STEM-MB-0104-CJ)

Introduction

Gymnodimines (GYMs) are the smallest members of cyclic imines (CIs) in terms of molecular weight. They are characterized as fast-acting toxins, which can accumulate in filter feeding shellfish and may pose a potential risk to human health and the aquaculture industry. Being first discovered in New Zealand oysters, GYMs have been detected in pipis (Donax deltoides), mussels (Modiolus proclivis), oysters (Saccostrea glomerata), and seawater in many countries. GYMs can also be isolated from the dinoflagellates Alexandrium ostenfeldii (Alexandrium peruvianum ) and Karenia selliformis, which are harmful algal bloom species. The stereochemical structure of gymnodimine-A (GYM-A) was first elucidated by nuclear magnetic resonance (NMR) in 1995. It respective molecular weight is around 520. As liposoluble toxins, the typical chemical structure of GYMs contains a macrocycle ring, a spirocyclic imine ring, and an ether moiety.

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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; Toxicology

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: Buffer

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