title: Fluorescent Probe Encapsulated in Avidin Protein to Eliminate Nonspecific Fluorescence and Increase Detection Sensitivity in Blood Serum.
abstract: Quantitative detection of trace amounts of a biomarker in protein rich human blood plasma using fluorescent probes is a great challenge as the real signal is usually obscured by nonspecific fluorescence. This problem occurs because most of the fluorescent dyes bind very tightly with blood proteins to produce a large fluorescence increase, resulting in overestimation of the biomarker concentrations and false positive diagnosis. In this paper, we report that biotinylated fluorescent probes encapsulated in avidin protein can generate very specific fluorescence in blood serum by blocking out nonspecific dye-protein interactions. We applied our novel probe design to detect two different types of biomolecules, hydrogen sulfide and nitroreductase. Our Avidin conjugated probes achieved quantitative analyte detection in blood serum; whereas concentrations were overestimated up to 320-fold when bare fluorescent probes were employed. As compared to conventional approaches where fluorescent probes are encapsulated into polymers and nanoparticles, our simple approach successfully overcomes many key issues such as dye leakage, long preparation steps, inconsistent dye-host ratios, difficulty in constructing in situ in a complex medium, and limited application to detect only small metabolites.
authors.affiliation: Department of Chemistry, National Tsing Hua University , 101 Section 2, Kuang Fu Road, Hsinchu 30013, Taiwan, Republic of China.
authors.forename: Ting-Wei
authors.initials: TW
authors.lastname: Wu
authors.affiliation: Department of Chemistry, National Tsing Hua University , 101 Section 2, Kuang Fu Road, Hsinchu 30013, Taiwan, Republic of China.
authors.forename: Fang-Hong
authors.initials: FH
authors.lastname: Lee
authors.affiliation: Department of Chemistry, National Tsing Hua University , 101 Section 2, Kuang Fu Road, Hsinchu 30013, Taiwan, Republic of China.
authors.forename: Ruo-Cing
authors.initials: RC
authors.lastname: Gao
authors.affiliation: Department of Chemistry, National Tsing Hua University , 101 Section 2, Kuang Fu Road, Hsinchu 30013, Taiwan, Republic of China.
authors.forename: Chee Ying
authors.initials: CY
authors.lastname: Chew
authors.affiliation: Department of Chemistry, National Tsing Hua University , 101 Section 2, Kuang Fu Road, Hsinchu 30013, Taiwan, Republic of China.|Frontier Research Center on Fundamental and Applied Sciences of Matters, National Tsing Hua University , 101 Section 2, Kuang Fu Road, Hsinchu 30013, Taiwan, Republic of China.
authors.forename: Kui-Thong
authors.initials: KT
authors.lastname: Tan
mesh_terms: D001360:Avidin / Q000097:blood / Q000737:chemistry*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D006862:Hydrogen Sulfide / Q000097:blood*; D009601:Nitroreductases / Q000097:blood* / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D001360:Avidin; D009601:Nitroreductases; D006862:Hydrogen Sulfide