Compositions for the detection of enzyme activity in...

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/37 (2006.01) A61K 38/00 (2006.01) G01N 21/00 (2006.01) G01N 21/76 (2006.01) G01N 33/533 (2006.01)

Patent

CA 2384021

The present invention provides for novel reagents whose fluorescence increases in the presence of particular proteases. The reagents comprise a characteristically folded peptide backbone conjugated to two fluorophores such that the fluorophores are located opposite sides of a cleavage site. When the folded peptide is cleaved, as by digestion with a protease, the fluorophores provide a high intensity fluorescent signal at a visible wavelength. Because of their high fluorescence signal in the visible wavelengths, these protease indicators are particularly well suited for detection of protease activity in biological samples, in particular in frozen tissue sections. Thus this invention also provides for methods of detecting protease activity in situ in frozen sections.

La présente invention porte sur de nouveaux réactifs dont la fluorescence augmente en présence de protéases spécifiques. Ces réactifs comprennent un squelette de peptide replié de manière caractéristique et conjugué à deux fluorophores de sorte que les fluorophores soient situés sur les côtés opposés d'un site de clivage. Lorsque le peptide plié est clivé, par la digestion avec une protéase, les fluorophores génèrent un signal fluorescent de haute intensité à une longueur d'onde visible. Du fait de leur haute fluorescence dans les longueurs d'onde visibles, ces indicateurs de protéases sont notamment appropriés pour détecter l'activité des protéases dans des échantillons biologiques, notamment dans des sections de tissu gelées. Cette invention porte également sur des procédés de détection de l'activité des proétases in situ dans des sections de tissu gelées.

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