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Quantum dots (ODs) with unique optical properties such as size-tunable emission , simultaneous excitation, high brightness, and very slow photobleaching ¿can be used as powerful cancer diagnostic tools providing the -"'¿cile-cular¿ profiles of cancer cases based on common clinical biopsies and allowing the tumor cell detection ¿and-analysis in highly heterogenous samples and rare cell populations. However, researchers have found that the¿ fluorescence intensity of QDs changes under light irradiation and the QD-antibody conjugates detaches from tissues easily during the staining process, making it very difficult to quantify expression level of biomarkers. To develop QD based multiplexing and quantitative kits for clinical applications, this team proposes to increase the QD stability through the multi-shell coating strategy in Aim 1 and to develop the high-bioaffinity QD-Antibody (Ab) through the adaptor protein strategy in Aim 2. As the first application, the improved QO-Ab technology will be used to develop a multi-biomarker imaging kit for prediction of human papilomavirus (HPV)- associated cancer. HPV is one of the risk factors in cancer development. Accurate evaluation of the impact of HPV on cancer development requires incorporation of our knowledge of the pathways by which HPV causes cancer. This team would like to use three QD-conjugated antibodies which specifically bind to HPV-related proteins as a predictor for cancer assessment. As a preparation for the phase II development of this technology, Aim 3 and 4 will validate Ihe HPV specific QD-Abs and oplimize Ihe condition for multiple QD-staining and quantification, respectively, by using human cancer cell lines and tissues.