METHOD FOR DETECTION OF SPECIFIC IMMUNOGLOBULIN CLASS G ANTIBODIES

• US 20110143454A1
• Filed: 06/17/2010
• Published: 06/16/2011
• Est. Priority Date: 05/08/2007
• Status: Active Grant

• Current Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Original Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Inventors: Upmeier, Barbara, Bollhagen, Ralf, Bronold, Martina, Markert-Hahn, Christine, Donie, Frederic
• Granted Patent: US 9,267,953 B2
• Time in Patent Office: Days
• US Class Current: 436/501
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/6854 Immunoglobulins

Abstract

The invention relates to a method for determining an analyte in a sample by means of an assay that can be carried out in a one-step format without performing washing steps. The method includes a first analyte-specific receptor that contains at least one bonding point for the analyte, e.g., several antibodies on a particle, as well as a second analyte-specific receptor that can selectively bond to an arrangement comprising at least two analyte molecules which are bound to the first receptor. An antibody sandwich immune assay is described in which the detection is based on electrochemiluminescence.

Claims

1. A method for determining an analyte in a sample comprising the steps of:

contacting the sample with a first analyte-specific receptor and a second analyte-specific receptor wherein one of the two receptors is bound to a solid phase or is capable of binding to a solid phase and the other receptor carries a signal-generating group or is capable of binding to a signal-generating group, and

detecting the presence or/and the amount of the analyte by determining the signal-generating group on the solid phase,

wherein the first analyte-specific receptor contains at least two binding sites for the analyte and the second analyte-specific receptor is capable of selectively binding to an arrangement of at least two analyte molecules which are bound to the first receptor, wherein the at least two analyte molecules bound to the first receptor are aggregated.

2. The method according to claim 1 wherein the analyte is an antigen-specific antibody of the immunoglobulin class G.

3. The method according to claim 1 wherein the first receptor is bound to the solid phase or is capable of binding to the solid phase and the second receptor carries the signal-generating group or is capable of binding to the signal-generating group.

4. The method according to claim 1 wherein the second receptor is bound to the solid phase or is capable of binding to the solid phase and the first receptor carries the signal-generating group or is capable of binding to the signal-generating group.

5. The method according to claim 1 wherein the first receptor contains at least two different binding sites, at least two identical binding sites or a combination of different and identical binding sites for the analyte.

6. The method according to claim 1 wherein the first receptor is an oligomeric or a multimeric receptor.

7. The method according to claim 1 wherein the first receptor comprises an antigen which contains at least two epitopes as binding sites for the analyte.

8. The method according to claim 1 wherein the first receptor comprises a bead.

9. The method according to claim 1 wherein the first receptor is provided in the form of receptor precursors.

10. The method according to claim 9, wherein the receptor precursors are of such a nature that they can associate with one another or bind to one another during or before the contacting step in order to form the first receptor.

11. The method according to claim 1 wherein the second receptor is a rheumatoid factor-like antibody.

12. The method according to claim 1 wherein the second receptor is an antibody with at least two paratopes.

13. The method according to claim 1 wherein the second receptor is an antibody of the immunoglobulin class M.

14. The method according to claim 1 wherein the second receptor is an antibody having low affinity and high avidity for the analyte.

15. The method according to claim 1 wherein the second receptor is a monoclonal antibody.

16. The method according to claim 1 wherein the contacting step is carried out in one reaction mixture without a washing step.

17. The method according to claim 1 which is a diagnostic method.

18. A kit comprising a first analyte-specific receptor and a second analyte-specific receptor, wherein one of the two receptors is bound to a solid phase or is capable of binding to a solid phase and the other receptor carries a signal-generating group or is capable of binding to a signal-generating group, wherein the first analyte-specific receptor contains at least two binding sites for the analyte and the second analyte-specific receptor is capable of selectively binding to an arrangement of at least two analyte molecules which are bound to the first receptor, wherein the at least two analyte molecules bound to the first receptor are aggregated.

19. The kit according to claim 18, wherein the first analyte-specific receptor and the second analyte-specific receptor are formulated for use in one reaction mixture.

20. The kit according to claim 18 wherein the first receptor is bound to the solid phase or is capable of binding to the solid phase and the second receptor carries the signal-generating group or is capable of binding to the signal-generating group.

21. The kit according to claim 18 wherein the second receptor is bound to the solid phase or is capable of binding to the solid phase and the first receptor carries the signal-generating group or is capable of binding to the signal-generating group.