Label free detection of biologically relevant binding pairs has provided critical insight into the characterization of reagents used in both therapeutic and diagnostic applications. The Interferometric Reflectance Imaging Sensor (IRIS) platform has been developed for the multiplexed, real-time detection of such binding interactions. Improvements to experimental methodology and analysis applied to the latest iteration of the IRIS provided heretofore unseen binding characterizations with this multiplexed platform. Here, we extend and demonstrate the utility of the IRIS system to (1) evaluate and compare kinetic parameters to those obtained with more traditional label free methods (2) characterize multiple, disease relevant antibodies in multiple disease systems (anthrax, Zika, dengue and plague) (3) determine appropriate binding pairs in multiplexed label free formats and (4) obtain 10-fold improvements to the limits of detection for analyte in solution over previous IRIS iterations. Applications to immunoassay development are discussed throughout with exemplary datasets provided. Observations regarding additional IRIS utilities are also discussed, including qualifications of genetically engineered ligands, evaluating subcloned antibodies and screening unpurified antibody supernatants.
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/36604 |
| Date | 13 June 2019 |
| Creators | Needham, James William |
| Contributors | Ünlü, M. Selim |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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