Nucleic acid probe linked to a label having a terminal cysteine

• US 5,109,124 A
• Filed: 02/02/1989
• Issued: 04/28/1992
• Est. Priority Date: 06/01/1988
• Status: Expired due to Fees

• Current Assignee: Biogen, Inc.
• Original Assignee: Biogen, Inc.
• Inventors: Cate, Richard L., Ramachandran, Kuzhalmannam L.
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Marschel, Ardin H.
• Application Number:
  US07/306,798
• Time in Patent Office: 1,181 Days
• Field of Search: 435/6, 435/91, 436/501, 536/27, 935/19, 935/78, 935/86, 935/88
• US Class Current: 536/24.3
• CPC Class Codes: A61K 2121/00 Preparations for use in therapy
• CPC Class Codes: A61K 51/088 conjugates with carriers being peptides, polyamino acids or proteins antibodies A61K51/10
• CPC Class Codes: A61K 51/1093 conjugates with carriers being antibodies
• 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

Abstract

A polynucleotide probe with a label bearing a plurality of signalling moieties. The label is attached to the probe by the reaction of an amino and sulfhydryl reactive hetero bifunctional reagent with the probe and label, the reaction resulting in the oxidation of a sulfhydryl group of the label. The label may be attached to the 5'"'"' terminus of the probe, or to modified bases of the probe. Probes constructed according to the invention are useful in detecting target sequences in genomic DNA.

Claims

1. A labeled probe comprising a polynucleotide sequence complementary to a target polynucleotide sequence, a label, including aterminal cysteine residue, and a linkage group, the label and linkage group not interfering substantially with the characteristic ability of the probe polynucleotide sequence to hybridize to the target polynucleotide sequence, the label being covalently attached to the linkage group through a sulfur-carbon single bond, the sulfur atom of which is part of the terminal cysteine residue of the label, and the linkage group being covalently attached to the polynucleotide sequence through a carbon-oxygen single bond.

2. The labeled probe of claim 1, wherein the label comprises a polypeptide, including a terminal cysteine residue.

3. The labeled probe of claim 1, wherein at least one signalling moiety is attached to the label.

4. The labeled probe of claim 3, wherein at least one signalling moiety is attached to the label by way of at least one bridging moiety.

5. The labeled probe of claim 4, wherein the bridging moiety is selected from the group consisting of biotin, iminobiotin, avidin, streptavidin and an antibody to the label.

6. The labeled probe of claim 3, 4 or 5, wherein the signalling moiety is selected from the group consisting of radioactive isotopes, enzymes, coumarin, and fluorescent compounds.

7. The labeled probe of claim 1, 2, 3 or 4 wherein the linkage group is attached at the 5'"'"' terminus of the polynucleotide.