Single-molecule phosphorescence immunoassay microscopy was developed and applied for high-throughput screening of tumor markers at the single-molecule (SM) level with no need of separation processes. The screening of individual analyte in a mixture was based upon distinguished diffusion images of single molecules associated with its size and mass. As a working example, several molecular forms of serum prostate- specific antigens (PSA) were labeled with Ru(bpy)32+-NHS-ester and labeled PSA-free and PSA-complex were distinguished based upon their SM diffusion images using this SM microscopy. The bound and unbound PSA-free with its monoclonal antibody (MAB) were also detected using this SM microscopy. A novel solution-phase quantitative electro chemiluminescence (ECL) immunoassay was developed to measure affinity constants of PSA with its antibody and diffusion coefficients of labeled PSA. The ECL immunoassay was able to detect PSA at 1.7 pg/mL. Diffusion of labeled PSA-free and PSA-complex measured by ECL and SM microscopy was consistent demonstrating that distinguished SM diffusion images could be used to screen multiple analytes in a complex mixture. This also implied the possibility of real- time monitoring of kinetics of binding reactions using such SM microscopy.
Conference Committee Involvement (6)
Advanced Biomedical and Clinical Diagnostic Systems VI
20 January 2008 | San Jose, California, United States
Advanced Biomedical and Clinical Diagnostic Systems V
21 January 2007 | San Jose, California, United States
Advanced Biomedical and Clinical Diagnostic Systems IV
22 January 2006 | San Jose, California, United States
Advanced Biomedical and Clinical Diagnostic Systems III
23 January 2005 | San Jose, CA, United States
Advanced Biomedical and Clinical Diagnostic Systems
25 January 2004 | San Jose, CA, United States
Advanced Biomedical and Clinical Diagnostic Systems
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