Fluorescent dyes and uses thereof
The present invention provides fluorescent dyes that are based on firefly luciferin structure. These dyes are optimally excited at shorter wavelengths and have Stokes shift of at least 50 nm. The fluorescent dyes of the invention are useful for preparation of dye-conjugates, which can be used in detection of an analyte in a sample.
1. A dye-conjugate having the formula:
wherein
X 1 and X 2 are independently S or O;
Y 1 and Y 2 separately represent halogen, or Y 1 is CF 3 and Y 2 is H;
Z 1 is H or alkyl;
wherein Z 1 and one of the W groups may be absent and, if absent, are replaced with an additional bond between C 4 and C 5 in ring C;
L is selected from the group consisting of a carbonyl group (C═O), —C(═O)NH(CH 2 ) n C(O)— wherein n is an integer from 1-5, and —(CH 2 ) n C(═O)— wherein n is an integer from 1-5; and
wherein CM is a covalently linked conjugated molecule comprising a protein.
2. The dye-conjugate of claim 1 wherein the protein is an antibody, or a fragment of an antibody.
3. The dye-conjugate of claim 2 wherein the antibody is selected from the group consisting of an anti CD3 antibody, an anti CD4 antibody, and an anti CD45 antibody.
4. The dye-conjugate of claim 1 wherein Y 1 and Y 2 are each a halogen.
5. The dye-conjugate of claim 1 wherein Y 1 and Y 2 are each chlorine.
6. The dye-conjugate of claim 1 wherein Y 1 and Y 2 are each fluorine.
7. The dye-conjugate of claim 1 wherein Y 1 is CF 3 and Y 2 is H.
8. The dye-conjugate of claim 1 wherein each W is alkyl.
9. The dye-conjugate of claim 1 wherein each W is methyl.
10. The dye-conjugate of claim 1 wherein more than one dye molecule is conjugated to the protein.
11. The dye-conjugate of claim 1 wherein X 1 and X 2 are both S atoms.
12. The dye-conjugate of claim 1 wherein X 1 is an S atom and X 2 is an O atom.