Synthesis And Application Of Novel Terbium Chelate-based Fluorescence Probes

Time-resolved fluorescence bioassay technique based on long-lived fluorescence of lanthanide probes is an ultrasenstive bioanalytical method, and has been widely used in clinical diagnostics and biotechnology. In this thesis, three types of Tb3+ chelate-based fluorescence probes, N,N,N1,N1-{2,6-bis(3'-aminomethyl-1'-pyrazolyl)-4-[N,N'-bis(2-picolyl )aminomethylene-pyridine]}tetrakis(acetate)-Tb3+(BBATA-Tb3+),N,N,N1,N1-[2,6-bis(3'-amio methyl-1'-pyrazolyl)-4-(4'-aminophenoxy)methyl-ene-pyridine]tetrakis(acetate)-Tb3+(BMPT A-Tb3+) and N,N,N1,N1-[2,6-(3'-aminomethyl-1'-pyrazolyl)-4-(3,4-diaminophenyl)-methoxyl pyridine]tetrakis(acetate)-Tb3+(BMTA-Tb3+) specific for Zn2+,·OH and H2O2 were designed, prepared, characterized and used for highly sensitive detection of Zn2+,-OH and H2O2, respectively.Three probes themselves are almost non-fluorescent, and the fluorescence intensity of BBATA-Tb3+, BMPTA-Tb3+ and BMTA-Tb3+ increased upon the addition of Zn2+,-OH and H2O2, respectively. The reaction of BMTA-Tb3+ with H2O2 was catalyzed by horseradish peroxidase (HRP). It is known that lanthanide(III) (mainly Eu3+ and Tb3+) complexes have very desirable features as luminescent probes, which make them be favorable for spatial determination of the intracellular concentration of an analyte through the time-gated measurement to minimize the interference from autofluorescence and light scatterings.In addition, it was found that the new probes could be easily transferred into living HeLa cells after esterified. A time-gated luminescence imaging technique that could fully eliminate the short-lived background fluorescence from cell components had been successfully developed for monitoring the Zn2+,·OH and H2O2 in living cells.