Methods for labeling a substrate using a hetero-diels-alder reaction

1. A method for selectively labeling a substrate, the method comprising:

providing a precursor compound having one of the formulas:

irradiating the precursor compound at a wavelength of 300 to 350 nm under conditions effective to form an o-naphthoquinone methide having one of the formulas:

and

contacting the o-naphthoquinone methide with a polarized olefin attached to a surface of a substrate and having the formula:

under conditions effective to form a hetero-Diels-Alder adduct,

wherein:

each R 1 is independently H, halogen, or an organic group, with the proviso that at least one R 1 comprises a detectable label;

each R 2 is independently H or an organic group;

each R 3 is independently H, halogen, or an organic group;

X is O or NR 4 ;

Y is OR 5 or NR 5 3 + (Z 1/q ) − wherein Z is an anion having a negative charge of q;

each R 5 is independently H or an organic group;

optionally, two or more R 1 groups may be combined to form one or more rings;

optionally, two or more R 2 groups may be combined to form one or more rings;

optionally, two or more R 3 and/or R 4 groups may be combined to form one or more rings;

optionally, two or more R 5 groups may be combined to form one or more rings; and

with the proviso that at least one R 2 , R 3 , or R 4 comprises a linker group attached to the surface of the substrate.

2. The method of claim 1 wherein the precursor compound is irradiated in the presence of the polarized olefin.

3. The method of claim 2 wherein irradiating the precursor compound comprises pattern-wise irradiating the substrate to provide a pattern-wise labeled surface of the substrate.

4. The method of claim 1 wherein conditions effective to form the o-quinone methide comprise irradiation of an aqueous solution, suspension, or dispersion of the precursor compound.

5. The method of claim 1 wherein the substrate is selected from the group consisting of glass, quartz, silica, a metal, a semi-conductor, a polymer, a membrane, a liposome, a micelle, a macromolecule, a biomaterial, and combinations thereof.

6. The method of claim 1 wherein the label is detectable by a method selected from the group consisting of fluorescence, phosphorescence, radiation detection, optical and electrochemical methods, surface plasmon resonance imaging (SPRi), and combinations thereof.

7. The method of claim 1 wherein the detectable label comprises a probe.

8. The method of claim 7 wherein the probe comprises DNA, a peptide, a polypeptide, a protein, or a combination thereof.

9. The method of claim 1 further comprising removing the detectable label from the surface of the substrate by hydrolyzing the C—X bond.

10. A method for selectively labeling a substrate, the method comprising:

providing a precursor compound having one of the formulas:

irradiating the precursor compound at a wavelength of 300 to 350 nm under conditions effective to form an o-naphthoquinone methide having one of the formulas:

and

contacting the o-naphthoquinone methide with a polarized olefin attached to a surface of a substrate and having the formula:

under conditions effective to form a hetero-Diels-Alder adduct,

wherein:

each R 1 is independently H, halogen, or an organic group, with the proviso that at least one R 1 comprises a detectable label;

each R 2 is independently H or an organic group;

each R 3 is independently H, halogen, or an organic group;

X is O or NR 4 ;

Y is OR 5 or NR 5 3 + (Z 1/q ) − wherein Z is an anion having a negative charge of q;

each R 5 is independently H or an organic group;

optionally, two or more R 1 groups may be combined to form one or more rings;

optionally, two or more R 2 groups may be combined to form one or more rings;

optionally, two or more R 3 and/or R 4 groups may be combined to form one or more rings;

optionally, two or more R 5 groups may be combined to form one or more rings;

with the proviso that at least one R 2 , R 3 , or R 4 comprises a linker group attached to the surface of the substrate, and

wherein the substrate comprises a planar surface or a bead.

11. The method of claim 5 wherein the biomaterial is selected from the group consisting of a virus, a small multicellular organism, DNA, RNA, a peptide, a polypeptide, a protein, a carbohydrate, a lipid, tissue, and combinations thereof.