Instrument design and optimization of interferometric reflectance imaging sensors for in vitro diagnostics
Reddington, Alexander P
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In the field of drug discovery and disease diagnostics, protein microarrays have generated much enthusiasm for their high-throughput monitoring of biomarkers; however, this technology has yet to translate from research laboratories to commercialization. The hindrance is the considerable uncertainty and skepticism regarding data obtained. The disparity in results from different laboratories performing identical tests is attributed to a lack of assay quality control. Unlike DNA microarrays, protein microarrays have a higher level of bioreceptor immobilization variability and non-specific binding because of the more complex molecular structure and broader physiochemical properties. Traditional assay detection modalities, such as fluorescence microscopy and surface plasmon resonance, are unable to overcome both of these sources of variation. This dissertation describes the hardware and software design and biological validation of three complementary platforms that overcome bioreceptor variability and non-specific binding for diagnostics. In order to quantify the bioreceptor quality, a label-free, nondestructive, low cost, and high-throughput interferometric sensor has been developed as a quality control tool. The quality control tool was combined with a wide-field fluorescence imaging system to improve fluorescence experimental repeatability. Lastly, a novel high-throughput and label-free platform for quality control and specific protein microarray detection is described. This platform overcomes the additional complexities and time required with labeled assays by discriminating between specific and nonspecific detection by including sizing of individual binding events. Protein microarrays may one day emerge as routine clinical laboratory tests; however, it is important that the proper quality control procedures are in place to minimize erroneous results. These platforms provide reliable and repeatable protein microarray measurements for new advancements in disease diagnostics with the potential for drug discovery.
Thesis (Ph.D.)--Boston University