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BioLayer Interferometry (BLI) is an analytical technique utilized for measuring binding interactions between biomolecules. This is a label-free technology that measures the change in white light signal as molecules, proteins, antibodies, or other ligands attach to the end of an optical sensor tip throughout an assay in real time. BLI is advantageous in detection because the sample preparation is free of tagging or other manipulations, samples can be analyzed without purification, and the sample is fully recoverable. BLI is commonly used to measure affinity between two or more molecules as a characterization technique, high throughput screening of molecules for bioactivity, and quantitation of molecules in reaction mixtures or cell lysates. We utilized BLI as a yes/no detection platform to replace prolonged immunoassays for biothreat surrogate detection and developed an accompanying data analysis tool to automate the decision-making process for inexperienced users. Proprietary anti-mouse capture biosensors bound to two separate monoclonal antibodies, which then bound varying concentrations of protein to measure the on- and off-rates of the protein to the antibody at equilibrium. Software fitting reports kinetics values that were used to develop a secondary screening tool to determine a true binding event over a false positive or nonspecific interaction between binding partners. Over 1300 binding curves were generated to define the parameters of this screening tool, resulting in high confidence in the yes/no decision process. Implementation of this tool reduces the expertise needed for biothreat detection in the field or a high-throughput screening scenario, while also reducing the time needed from sample receipt to answer 2-fold over ELISA or MAGPIX immunoassays.
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