Development of an Automated Continuous Flow System for Large-Scale Production of Highly Fluorescent and Functionalized Semiconductor Nanocrystals

In the Phase I work BioCrystal demonstrated the feasibility of producing highly fluorescent and functionalized semiconductor nanocrystals with a narrow size distribution in a systematic and reproducible manner, and further demonstrated that the propertiesof these fluorescent nanocrystals are superior to the properties of conventional organic dyes currently in use in fluorescence-based assays. BioCrystal further developed a novel surface chemistry that provides protection, stabilization (against oxidation,ionic strength and buffers), aqueous solubility in a wide range of pHs, functionalization and linking ability, and dramatic fluorescence enhancement (brightness).In Phase II BioCrystal will develop and successfully demonstrate a prototype automated continuous flow process system for industrial-scale production of high quality, functionalized semiconductor nanocrystals with narrow size distribution. The Phase IIeffort will also emphasize continuing research on techniques that are currently being developed in our laboratories for the functionalization of the nanocrystals for biological tagging. These nanocrystals will be directly conjugated to a variety ofbioreceptors (i.e., antibodies, avidin, nucleic acids, and lectins)that are commonly used in biological detection and diagnostics. These conjugated nanocrystals will be fully characterized and favorably compared in selected fluorescence-based assays withconjugates of organic fluorophores commonly used today.The Phase II effort will demonstrate BioCrystal's capability of producing industrial-scale levels of high quality, monodisperse, functionalized fluorescent semiconductor nanocrystals for broadindustrial applications in research and diagnostics. This effort will further demonstrate the efficacy of these functionalized nanocrystals for use in the following: nucleic acid diagnostics; DNA and antibody microarrays; immunohistochemistry; qualitativedisease detection schemes; for bio-agent detection, in either air-borne or in aqueous environments; in the many different fluorescence-based assays used for research in the life sciences and clinical medicine; and for non-medical applications inidentification and authentification.Upon successful demonstration of this industrial-scale production prototype system, BioCrystal will confirm its ability to adequately address market demand for fluorescent nanocrystals in the multi-billion dollar fluorescence-based bioreagent marketplace,either directly or through large, well-entrenched bioreagent company partners. BioCrystal will further demonstrate that it can readily provide functionalized fluorescent nanocrystal conjugates to the government for multiple applications in bio-detection.