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

摘要

Methods for labeling a substrate using a hetero-Diels-Alder reaction are disclosed. The hetero-Diels-Alder reaction includes the reaction of an o-quinone methide (e.g., an o-naphthoquinone methide) with a polarized olefin to form a hetero-Diels-Alder adduct. The o-quinone methide or the polarized olefin can be attached to a surface of a substrate, and the other of the o-quinone methide or the polarized olefin can include a detectable label. The o-quinone methide can conveniently be generated by irradiation of a precursor compound, preferably in an aqueous solution, suspension, or dispersion.

主权项

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 R1 is independently H, halogen, or an organic group, with the proviso that at least one R1 comprises a detectable label; each R2 is independently H or an organic group; each R3 is independently H, halogen, or an organic group; X is O or NR4; Y is OR5 or NR53+(Z1/q)− wherein Z is an anion having a negative charge of q; each R5 is independently H or an organic group; optionally, two or more R1 groups may be combined to form one or more rings; optionally, two or more R2 groups may be combined to form one or more rings; optionally, two or more R3 and/or R4 groups may be combined to form one or more rings; optionally, two or more R5 groups may be combined to form one or more rings; and with the proviso that at least one R2, R3, or R4 comprises a linker group attached to the surface of the substrate.