Fluorescent labelling compositions and methods for their use

• US 5,650,334 A
• Filed: 08/31/1995
• Issued: 07/22/1997
• Est. Priority Date: 08/31/1995
• Status: Expired due to Fees

• Current Assignee: First Medical, Inc.
• Original Assignee: First Medical, Inc.
• Inventors: Choo, Sae Hyun, Zuk, Robert F.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): NGUYEN, BAO THUY L
• Application Number:
  US08/521,860
• Time in Patent Office: 691 Days
• Field of Search: 435/7.1-7.21, 435/7.4, 435/7.5, 435/7.92, 435/7.93, 435/7.94, 435/40.52, 435/174, 435/960, 435/968, 436/508, 436/518, 436/519, 436/523, 436/524, 436/525, 436/529, 436/528, 436/530, 436/800
• US Class Current: 436/529
• CPC Class Codes: G01N 2474/20 Immunohistochemistry assay
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: Y10S 435/968 High energy substrates, e.g. fluorescent, chemiluminescent, radioactive
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine

Abstract

A fluorescent labelling composition comprises a linear polysaccharide backbone molecule having a plurality of target-binding molecules, such as antibodies or nucleic acids, attached at spaced-apart intervals thereon. Each of the target-binding molecules, in turn, includes a multiplicity of fluorescent dye molecules bound thereto. In this way, fluorescent signal introduced to a single target-site on a solid phase surface may be increased without loss of binding activity.

Claims

1. A method for labelling a target substance in or on a solid phase, said method comprising:

providing a labelling composition including;

(a) a linear polysaccharide having a molecular weight of at least 500 kD;

(b) a plurality of molecules which specifically bind to the target substance attached to spaced-apart locations along the linear polysaccharide, and

(c) a multiplicity of fluorescent dye molecules bound to at least some of the target-binding molecules; and

exposing the labelling composition to the solid phase.

2. A method as in claim 1, wherein the solid phase is selected from the group consisting of a polymeric surface, an inorganic surface, and a tissue section.

3. A method as in claim 2, wherein the labelling composition is exposed to a solid phase surface which has previously been prepared by:

providing a solid phase surface having a specific binding substance specific for the target substance immobilized thereon; and

exposing the solid phase to a biological sample suspected of containing the target substance, whereby target substance is bound to the solid phase; and

wherein the labelling composition exposing step comprises exposing the labelling composition to the solid phase.

4. A method as in claim 1, wherein the linear polysaccharide is a dextram.

5. A method as in claim 4, wherein the linear polysaccharide is dextrans having a molecular weight of at least 1000 kD.

6. A method as in claim 1, wherein the target binding molecules are selected from the group consisting of antibodies, antibody fragments, antibody analogs, antigens, nucleic acids, hormones, hormone receptors, enzymes, enzyme substrates, avidin, and streptavidin.

7. A method as in claim 1, wherein the linear polysaccharide is dextrans having a molecular weight of at least 1000 kD and the target binding molecule is an antibody.

8. A method as in claim 1, wherein the fluorescent dye molecules are cyanine dyes.

9. A method as in claim 8, wherein the cyanine dyes are arylsulfonate cyanines.

10. A method as in claim 9, wherein the arylsulfonate cyanine is Cy5.

11. A method as in claim 1, wherein each target-binding molecule has from 1 to 10 fluorescent dye molecules covalently bound thereto.

12. A method as in claim 1, wherein the linear polysaccharide is dextrans having a molecular weight of at least 1000 kD, the target binding molecule is an antibody, and the fluorescent dye is an arylsulfonate cyanine.