Luminescent dyes with a water-soluble intramolecular bridge and their biological conjugates
Chemically reactive dyes that are intramolecularly crosslinked with a water-soluble bridge, their bioconjugates and their uses are described. Reactive fluorescent dyes that have a water-soluble bridge are superior to those of conjugates of spectrally non-crosslinked dyes or the dyes that are crosslinked with a hydrophobic bridge. The invention includes reactive fluorescent dyes, their biological conjugates and uses.
1. A chemically reactive luminescent dye having the formula:
wherein L is a linker of 0-30 atoms length; and RM is an amine, a carboxylic acid, an activated ester of a carboxylic acid, a haloacetamide, a hydrazine, a hydroxylamine, an isothiocyanate, a maleimide, or a sulfonyl halide.
2. A luminescent dye conjugate having the formula:
wherein WSB is a water-soluble bridge comprising at least a sulfonate or a phosphonate group; BIOPOLYMER is a biological substance having a molecular weight of larger than 1000 daltons; n is an integer of 1-30 provided that the dye is covalently connected to the BIOPOLYMER directly or indirectly via a linker “L”; and L is a linker of 0-30 atoms in length; and
wherein said luminescent dye conjugate is derived from the reaction of:
a chemically reactive luminescent dye having the Formula:
wherein L is the linker of 0-30 atoms length, and RM is an amine, a carboxylic acid, an activated ester of a carboxylic acid, a haloacetamide, a hydrazine, a hydroxylamine, an isothiocyanate, a maleimide, or a sulfonyl halide; and
the BIOPOLYMER.
3. The conjugate of claim 2 wherein the BIOPOLYMER is a peptide, a protein, a polysaccharide, an oligonucleotide, a nucleic acid, a lipid, a phospholipid, a lipoprotein, a lipopolysaccharide, a liposome, a lipophilic polymer, a polymeric microparticle, an animal cell, a plant cell, a bacterium, a yeast, or a virus.
4. The conjugate of claim 2 wherein the BIOPOLYMER is an antibody.