Time-resolved luminescence binding assays using a fluorescent transition metal label other than ruthenium

• US 5,585,279 A
• Filed: 08/08/1994
• Issued: 12/17/1996
• Est. Priority Date: 01/23/1986
• Status: Expired due to Term

• Current Assignee: Lonza Group AG
• Original Assignee: Celltech Group Limited (UCB SA)
• Inventors: Davidson, Robert S.
• Primary Examiner(s): Green, Lora M.
• Application Number:
  US08/287,588
• Time in Patent Office: 862 Days
• Field of Search: 436/536, 436/537, 436/543, 436/544, 436/546, 436/84, 436/800, 436/805, 436/518, 436/172, 530/388.9, 530/389.8, 530/403
• US Class Current: 436/546
• CPC Class Codes: C09K 11/06 containing organic luminescent materials
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/553 Metal or metal coated
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/805 Optical property

Abstract

A time-resolved luminescence binding assay is described including a metal-labelled binding partner comprising a luminescent transition metal complex covalently attached to a binding partner such as an antigen or antibody. The metal-labelled binding partner, and a process for preparing said metal-labelled binding partner are described.

Claims

1. A time-resolved luminescence immunoassay method for detection of an analyte involving a metal-labelled binding partner, comprising the steps of:

a) binding the analyte to an immobilized anti-analyte first binding partner;

b) adding fluorescent metal-labelled second binding partner specific for the analyte, where the metal label comprises a luminescent transition metal complex, provided the transition metal is not ruthenium;

c) washing off the unbound labelled binding partner;

d) eluting the analyte-first binding partner-second fluorescent binding partner complex, and determining the fluorescence of the metal label by time-resolved fluorescence spectroscopy, thereby detecting the analyte.

2. The time-resolved luminescence immunoassay according to claim 1 wherein the transition metal is iridium, osmium or chromium.

3. The time-resolved luminescence immunoassay according to claim 1 wherein the luminescent transition metal complex comprises a transition metal complexed with one or more ligands, at least one of said ligands is provided with a chemical linkage covalently binding the luminescent transition metal complex to the binding partner.

4. The time-resolved luminescence immunoassay according to claim 1 wherein the luminescent transition metal complex comprises Ir(bipy)₂ diphenyl phenanthroline and the second binding partner comprises anti-TSH monoclonal antibody.

5. The time-resolved luminescence binding immunoassay method of claim 1 wherein the metal label comprises a luminescent transition metal complex provided that the metal label is not a ruthenium complex having the general formula:

space="preserve" listing-type="equation">Ru.sup.2+ L.sub.1 L.sub.2 L.sub.3

wherein L₁, L₂ and L₃ are the same or different and each is a bi- or polycyclic ligand with at least two nitrogen-containing heterocycles, at least one of these ligands is substituted with at least one group conferring water-solubility, and at least one of the ligands is substituted, optionally via a spacer group, with at least one reactive group, and the ligands L₁, L₂ and L₃ are attached to the ruthenium via nitrogen atoms.

6. The time-resolved luminescence binding immunoassay according to claim 5 wherein the luminescent transition metal complex comprises a transition metal complexed with one or more ligands, at least one of said ligands is provided with a chemical linkage covalently binding the luminescent transition metal complex to the binding partner.