Multisignal labeling reagents and processes and uses therefor

• US 9,884,889 B2
• Filed: 09/03/2015
• Issued: 02/06/2018
• Est. Priority Date: 04/03/2003
• Status: Expired due to Term

• Current Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Original Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Inventors: Coleman, Jack, Rabbani, Elazar, Stavrianopoulos, Jannis, Pande, Praveen
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US14/844,468
• Publication Number:
  US 20160025716A1
• Time in Patent Office: 887 Days
• Field of Search: 536 266
• CPC Class Codes: C07D 219/08 Nitrogen atoms
• CPC Class Codes: C07D 405/14 containing three or more hetero rings
• CPC Class Codes: C07D 491/22 in which the condensed system contains four or more hetero rings
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C09B 11/00 Diaryl- or thriarylmethane dyes
• CPC Class Codes: C09B 11/08 Phthaleins; Phenolphthaleins; Fluorescein
• CPC Class Codes: C09B 11/24 Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
• CPC Class Codes: C09B 57/02 Coumarine dyes
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

Provided are compounds comprising two DNA supramolecular binding molecules covalently joined by a linker group. Also provided are multisignal labeling reagents comprising (i) an oligomer of nucleotides or nucleotide analogs; (ii) a DNA supramolecular binding molecule noncovalently bound to the oligomer; and (iii) a first reactive group or a first partner of a first binding pair covalently bound to the oligomer. Additionally provided are methods of producing multisignal labeling reagents.

Claims

1. A multisignal labeling reagent comprising:

(i) an oligomer of nucleotides or nucleotide analogs;

(ii) a plurality of DNA supramolecular binding molecules noncovalently bound to the oligomer,

wherein said DNA supramolecular binding molecules comprise a detectable label attached thereto that is not a DNA supramolecular binding molecule, and

wherein said DNA supramolecular binding molecules are selected from minor groove binders and major groove binders; and

(iii) a first reactive group covalently bound to the oligomer, or

a first partner of a first binding pair covalently bound to the oligomer.

2. The multisignal labeling reagent of claim 1, wherein the DNA supramolecular binding molecule is fluorescent.

3. The multisignal labeling reagent of claim 1, wherein the detectable label is non-radioactive.

4. The multisignal labeling reagent of claim 3, wherein the non-radioactive detectable label is a fluorophore.

5. The multisignal labeling reagent of claim 3, wherein the non-radioactive detectable label is a chemiluminescent or phosphorescent moiety.

6. The multisignal labeling reagent of claim 1, wherein the DNA supramolecular binding molecules comprise DNA supramolecular binding molecules which are minor groove binders.

7. The multisignal labeling reagent of claim 6, wherein the minor groove binders comprise a minor groove binder selected from the group consisting of a diacrylamidine and a bis-benzimidazole.

8. The multisignal labeling reagent of claim 6, wherein the minor groove binders comprise a minor groove binder selected from the group consisting of DAPI, berenil, pentamidine, distamycin A, and Hoechst 33258.

9. The multisignal labeling reagent of claim 1, wherein the DNA supramolecular binding molecules comprise DNA supramolecular binding molecules which are major groove binders.

10. The multisignal labeling reagent of claim 9, wherein the major groove binders comprise a major groove binder selected from the group consisting of a zinc finger, a leucine zipper, and a helix-turn-helix motif.

11. The multisignal labeling reagent of claim 1, comprising the first partner of the first binding pair covalently bound to the oligomer.

12. The multisignal labeling reagent of claim 11, wherein the first binding pair is a ligand/receptor, a hormone/receptor, biotin/avidin, biotin/streptavidin or an antigen/antibody binding pair.

13. The multisignal labeling reagent of claim 11, wherein the first partner of the first binding pair is streptavidin.

14. The multisignal labeling reagent of claim 1, comprising the first reactive group covalently bound to the oligomer.