PMID: 11906256Mar 22, 2002Paper

A homogeneous DNA hybridization system by using a new luminescence terbium chelate

Bioconjugate Chemistry
Shinji SuedaKazuko Matsumoto

Abstract

Homogeneous DNA hybridization assay based on the luminescence resonance energy transfer (LRET) from a new luminescence terbium chelate, N,N,N(1),N(1)-[2,6-bis(3'-aminomethyl-1'-pyrazolyl)-4-phenylpyridine]tetrakis(acetic acid) (BPTA)-Tb(3+) (lambda(ex) = 325 nm and lambda(em) = 545 nm) to an organic dye, Cy3 (lambda(ex) = 548 nm and lambda(em) = 565 nm), has been developed. In the system, two DNA probes whose sequences are complementary to the two different consecutive sequences of a target DNA are used; one of the probes is labeled with the Tb(3+) chelate at the 3'-end, and the other is with Cy3 at the 5'-end. Labeling of the Tb(3+) chelate is accomplished via the linkage of a biotin-labeled DNA probe with the Tb(3+) chelate-labeled streptavidin. Strong sensitized emission of Cy3 was observed upon excitation of the Tb(3+) chelate at 325 nm, when the two probe DNAs were hybridized with the target DNA. The sensitivity of the assay was very high compared with those of the previous homogeneous-format assays using the conventional organic dyes; the detection limit of the present assay is about 30 pM of the target DNA strand.

Citations

Dec 17, 2005·Chemical Communications : Chem Comm·Rohan T Ranasinghe, Tom Brown
Jul 25, 2015·International Journal of Molecular Sciences·Alexander S GoryashchenkoAlexander P Savitsky
Jul 30, 2005·Chembiochem : a European Journal of Chemical Biology·Yoichiro KoshiItaru Hamachi
Mar 20, 2010·Chembiochem : a European Journal of Chemical Biology·Yulia V GerasimovaDmitry M Kolpashchikov
Sep 17, 2005·Journal of Fluorescence·Jingli Yuan, Guilan Wang
Feb 6, 2013·Biochemistry. Biokhimii︠a︡·V V Zherdeva, A P Savitsky
Dec 19, 2009·Analytical Chemistry·Ulla KarhunenTero Soukka
May 29, 2003·Journal of the American Chemical Society·Bin LiuGuillermo C Bazan
Jun 30, 2010·Chemical Reviews·Dmitry M Kolpashchikov
Sep 2, 2004·Journal of the American Chemical Society·Huixiang Li, Lewis J Rothberg

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