Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay

The overall goal of this bio-layer interferometry or BLI assay, is to measure the potential interaction between purified hEAG1 ion channel protein and small molecule lipids. This method can help answer key questions in the ion channel pharmacology field, such as whether the small molecule of interest can directly interact with this ion channel and what are the kinetics of the interaction? The main advantage of this technique is that it is a labor free and high throughput method.

Only a small amount of immobilized protein is needed. Generally, individuals who are new to this method will struggle because a successful assay involves multiple steps, including protein expression, purification, labeling, accurately tuned sensor, and data analysis. After culturing HEK293 T cells stabling expressing hEAG1 for two to three days, harvest the cells by removing the growth medium, and use four milliliters of PBS to wash the cells twice.