Non-radioactive nucleic acid hybridization probes

• US 4,996,142 A
• Filed: 09/04/1987
• Issued: 02/26/1991
• Est. Priority Date: 09/04/1986
• Status: Expired due to Fees

• Current Assignee: Agricultural Genetics Co. Ltd. (BASF SE)
• Original Assignee: Agricultural Genetics Co. Ltd. (BASF SE)
• Inventors: Hull, Roger, Al-Hakim, Ali H.
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Marschel, Ardin H.
• Application Number:
  US07/094,133
• Time in Patent Office: 1,271 Days
• Field of Search: 435/6, 530/358, 530/401, 521/31, 526/262, 536/27, 536/28, 536/29, 935/10, 935/77, 548/113, 548/181, 549/32, 549/50
• US Class Current: 435/6.12
• 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

The invention relates to an adduct of a basic macromoleculer and biotin having the general formula: ##STR1## in which MM is the residue of a basic macromolecule formed by substituting the H-atom of one or more primary amino groups;

alk represents an optionally substituted alkylene group having from 1 to 10 C-atoms;

m is 0 or 1; and

n is a positive integer,

with the proviso that, where MM represents cytochrome C or histone, m is not 0.

The adduct is useful in constructing non-radioactive nucleic acid hydridization probes.

Claims

1. An adduct of a basic macromolecule and biotin having the formula:

##STR5## wherein MM is the residue of a basic macromolecule selected from the group consisting of cytochrome C, histone and polyethyleneimine, formed by substituted the H-atom of one or more primary or secondary amino groups;

alk represents an alkylene group having from 1 to 10 C-atoms;

m is 0 or 1, and n is a positive integer from 1 to 30, with the proviso that, where MM represents cytochrome C or histone, m is not zero.

2. The adduct according to claim 1, wherein macromolecule MM is cytochrome C or histone.

3. The adduct according to claim 1, wherein macromolecule MM is polyethyleneimine (PEI).

4. A biotin-labelled nucleic acid, comprising:

the adduct of claim 1 linked to a nucleic acid by a cross-linking agent selected from the group consisting of formaldehyde, glutaraldehyde, 1, 2, 7, 8-diepoxyoctane, bis(succinimidyl) suberate and bis(sulphosuccinimidyl) suberate.

5. An adduct according to claim 3, in which the PEI has a molecular weight of from 1000 to 2000.

6. An adduct according to claim 1, in which alk is (--CH₂ --)₅.