Glycoconjugated fluorescent labeling reagents

• US 5,877,310 A
• Filed: 04/25/1997
• Issued: 03/02/1999
• Est. Priority Date: 04/25/1997
• Status: Expired due to Fees

• Current Assignee: Carnegie Mellon University
• Original Assignee: Carnegie Mellon University
• Inventors: Reddington, Mark V., Waggoner, Alan S.
• Primary Examiner(s): Higel, Floyd D.
• Application Number:
  US08/847,482
• Time in Patent Office: 676 Days
• Field of Search: 536/6.5, 536/25.32, 530/802, 435/7.8, 548/455
• US Class Current: 536/25.32
• CPC Class Codes: C07H 17/02 Heterocyclic radicals containing only nitrogen as ring hetero atoms
• CPC Class Codes: C08B 37/0012 Cyclodextrin [CD], e.g. cycle with 6 units (alpha), with 7 units (beta) and with 8 units (gamma), large-ring cyclodextrin or cycloamylose with 9 units or more; Derivatives thereof
• CPC Class Codes: C08B 37/0021 Dextran, i.e. (alpha-1,4)-D-glucan; Derivatives thereof, e.g. Sephadex, i.e. crosslinked dextran
• CPC Class Codes: C09B 23/02 the polymethine chain containing an odd number of >CH- or >C[alkyl]- groups
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/58 involving labelled substances G01N33/53 takes precedence
• CPC Class Codes: Y10S 530/802 Chromogenic or luminescent peptides

Abstract

Water soluble glycoconjugated fluorescent labeling reagents comprised of a fluorescent dye, one or more sugar or carbohydrate residues that impart water solubility, and a reactive group that allows covalent attachment of the reagent to a substrate are disclosed. Glycoconjugated fluorescent labeling reagents are depicted as follows: ##STR1## These glycoconjugates are highly water soluble because of the sugar residues. This property is expected to lower nonspecific binding to cellular matter, reduce precipitation of labeled substrates, inhibit quenching of fluorescence, and unlike many prior fluorescent labeling reagents, that are soluble only in organic solvents, render the glycoconjugates usable in wholly aqueous solution.

Claims

1. A method for imparting fluorescent properties to a target material, the method comprising the steps of incubating:

(i) a target material having at least one functional group selected from the group consisting of carboxylic acid, alkyl halide, aldehyde, ketone, amino, or sulphydryl or having at least one reactive group that can covalently bond with said at least one functional group, and;

(ii) an amount of a glycoconjugate fluorescent compound having the formula;

##STR3## wherein dye is a fluorophone;

sugar is a sugar or modified carbohydrate rendering the glycoconjugate soluble in aqueous media; and

Reactive group is a functional group selected from the group consisting of succinimidyl ester, carboxylic acid, isothiocyanate, haloacetamide, maleimide, alkyl halide, azido, hydrazido, aldehyde, ketone, amino sulphydryl, provided said reactive group can covalently bond with said at least one functional group;

for a period of time sufficient to permit said at least one functional or reactive group of said fluorescent compound to covalently bond to said at least one reactive or functional group of said target material.