Labeling reagents and methods of their use
View Patent ↗The present disclosure is directed to a reactive ester agent capable of conjugating a reporter molecule to a carrier molecule or solid support. The reactive ester agent has the general formula: wherein the variables are described throughout the application.
1. A method of making a compound of Formula I comprising:
contacting a carrier molecule or a solid support comprising a nucleophile with a compound of Formula IA or a tautomer or salt thereof:
wherein,
L is a linker;
R 1 is chlorine;
R 2 is chlorine;
R 3 is COO − , SO3 − , substituted azenyl, PEG, phosphate or bisphosphonate; and
R a is a reporter molecule;
forming a compound of Formula I or a tautomer or salt thereof:
wherein
L is the linker;
R a is the reporter molecule; and
R b is the carrier molecule or solid support comprising a nucleophile (X).
2. The method of claim 1 , wherein X is —NH, —S— or —O—.
3. The method of claim 1 , wherein L is a covalent bond, -alkyl-, -substituted alkyl-, -alkenyl-, -substituted alkenyl-, -heterocyclyl-, -substituted heterocyclyl-, -aryl-, -substituted aryl-, -heteroaryl-, -substituted heteroaryl-, -cycloakyl-, -substituted cycloalkyl-, -oxy-, -alkoxy-, -substituted alkoxy-, -thio-, -amino-, or -substituted amino-.
4. The method of claim 1 , wherein R a is a dye.
5. The method of claim 4 , wherein the dye is a xanthene, a cyanine, an indole, a benzofuran, a coumarin, or a borapolyazaindacine.
6. The method of claim 1 , wherein the reporter molecule R a is an ion chelating moiety, a hapten, an antibody, an enzyme, a radiolabel, a metal ion or metal ion containing substance, a pigment, a chromogen, a phosphor, a fluorogen, a bioluminescent substance, a chemiluminescent substance, or a semiconductor nanocrystal.
7. The method of claim 1 , wherein R b is a solid support.
8. The method of claim 1 , wherein R b is a carrier molecule.
9. The method of claim 8 , wherein the carrier molecule is selected from the group consisting of an amino acid, a peptide, a protein, a carbohydrate, a polysaccharide, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid polymer, a drug, a lipid, and a synthetic polymer.
10. The method of claim 1 , wherein the compound of Formula IA is a salt.
11. The method of claim 10 , wherein the salt comprises a potassium or sodium ion.
12. The method of claim 1 , further comprising incubating the carrier molecule or solid support with the compound of Formula IA after the contacting step.
13. The method of claim 1 , wherein the contacting step is done in an aqueous solution.
14. The method of claim 1 , wherein the reporter molecule is hydrophobic.
15. The method of claim 1 , wherein the compound of Formula IA is soluble in an aqueous solution.
16. A method of labeling a carrier molecule or solid support comprising:
contacting the carrier molecule or solid support with a compound of Formula IA or a tautomer or salt thereof:
wherein,
L is a linker;
R 1 is chlorine;
R 2 is chlorine;
R 3 is COO − , SO3 − , substituted azenyl, PEG, phosphate or bisphosphonate; and
R a is a reporter molecule; and
the carrier molecule or solid support comprises a nucleophile; and
forming a labeled carrier molecule or solid support.
17. The method of claim 16 , wherein the labeled carrier molecule or solid support comprises a compound of Formula I:
wherein,
R a is the reporter molecule; and
R b is the carrier molecule or solid support comprising a nucleophile (x).