Tropinin I calibrator and method of use thereof in a sandwich immunoassay

• US 5,925,533 A
• Filed: 12/13/1996
• Issued: 07/20/1999
• Est. Priority Date: 12/21/1995
• Status: Expired due to Term

• Current Assignee: Siemens Healthcare Diagnostics GmbH (Siemens AG)
• Original Assignee: Bayer AG
• Inventors: Petesch, Nicole, Petry, Christoph, Doth, Margit
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/763,374
• Time in Patent Office: 949 Days
• Field of Search: 435/7.1, 435/7.92, 435/7.93, 435/7.94, 435/7.95, 435/70.21, 435/240.27, 435/965, 435/967, 435/972, 436/518, 436/531, 436/8, 436/811, 436/815, 436/15, 530/391.1, 530/391.5, 530/391.7, 530/391.9, 530/807
• US Class Current: 435/7.94
• CPC Class Codes: G01N 33/531 Production of immunochemical test materials
• CPC Class Codes: G01N 33/6854 Immunoglobulins
• CPC Class Codes: G01N 33/96 involving blood or serum control standard
• CPC Class Codes: Y10S 435/967 Standards, controls, materials, e.g. validation studies, buffer systems
• CPC Class Codes: Y10S 435/972 Modified antibody, e.g. hybrid, bifunctional
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function
• CPC Class Codes: Y10S 436/815 Test for named compound or class of compounds
• CPC Class Codes: Y10S 530/807 Hapten conjugated with peptide or protein
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/105831 Protein or peptide standard or control [e.g., hemoglobin, etc.]

Abstract

A conjugate consisting of a sequence of the analyte and an antibody against one of the antibodies used in the test can be employed, in aqueous solution and in precisely known quantity, as a stable calibrator in a sandwich immunoassay for detecting the analyte.

Claims

1. A calibrator for use in a sandwich immunoassay for tropinin I, wherein the tropinin I is bound between a first antibody or fragment thereof and a labelled second antibody or fragment thereof said calibrator comprising (a) a third antibody or fragment thereof which specifically binds either (i) to said first antibody or fragment thereof or (ii) to said labelled second antibody or fragment thereof, said third antibody or fragment thereof conjugated to (ii) at least one peptide which consists of an antibody binding site of said troponin I and which specifically binds to whichever of either (i) said first antibody or fragment thereof or (ii) said labelled second antibody or fragment thereof which is not bound by said third antibody or fragment thereof.

2. The calibrator according to claim 1 wherein the at least one peptide has a length of from 4 to 30 amino acids.

3. The calibrator according to claim 1 wherein said third antibody or fragment thereof is conjugated to 1 to 50 peptides, and wherein the peptides consist of 1 to 20 different amino acid sequences of said troponin I.

4. The calibrator according to claim 1 wherein said calibrator is in an aqueous solution.

5. The calibrator according to claim 1 wherein the at least one peptide has an amino acid sequence selected from the group consisting of SEQ ID NO.:

1 and SEQ ID NO.;

2.

6. The calibrator according to claim 1 further comprising one or more auxiliary substances selected from the group consisting of a buffer, a stabilizer, a preservative, a detergent and a cosolvent.

7. In a sandwich immunoassy for determining tropinin I in a sample, wherein the tropinin I in the sample is bound between a first antibody or fragment thereof and a labelled second antibody or fragment thereof to form a complex and the amount of formed complex is measured relative to the amount of complex formed by a known amount of troponin I calibrator to determine the amount of the tropinin I in the sample, wherein the improvement comprises using as the calibrator

a calibrator comprising (a) a third antibody or fragment thereof which specifically binds either (i) to said first antibody or fragment thereof or (ii) to said labelled second antibody or fragment thereof, said third antibody or fragment thereof conjugated to (ii) at least one peptide which consists of an antibody binding site of said troponin I and which specifically binds to whichever of either (i) said first antibody or fragment thereof or (ii) said labelled second antibody or fragment thereof which is not bound by said third antibody or fragment thereof.