Process and reagent for the determination of a reaction partner of an immunological reaction

• US 4,880,731 A
• Filed: 03/16/1987
• Issued: 11/14/1989
• Est. Priority Date: 03/19/1986
• Status: Expired due to Term

• Current Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Original Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Inventors: Kaspar, Klaus P.
• Primary Examiner(s): NOT, DEFINED
• Assistant Examiner(s): Griffith, Jr., D. John
• Application Number:
  US07/025,876
• Time in Patent Office: 974 Days
• Field of Search: 436/531, 436/532, 436/533, 436/518, 436/523, 436/512, 436/809, 436/819, 436/825, 436/826, 436/540, 435/7
• US Class Current: 435/7.94
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: Y10S 435/962 Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/819 Multifunctional antigen or antibody
• CPC Class Codes: Y10S 436/825 Pretreatment for removal of interfering factors from sample
• CPC Class Codes: Y10S 436/826 Additives, e.g. buffers, diluents, preservatives

Abstract

For the determination of a reaction partner of an immunological reaction according to the principle of immunoassay, the reaction partner to be determined is brought into contact with a marked specific receptor R₁ and at least 1 unmarked receptor R₂, one of the unmarked receptors R₂ being bonded on a solid phase by a binding-capable substance R₃. In order to determine the sample blank value, the unmarked receptor R₂, which is bonded by the receptor fixed on the solid phase, is then replaced by another unmarked receptor R'"'"'₂ which does not react with the reaction partner of R₂ to be determined.

Claims

1. In an immunoassay for the determination of an immunological reaction partner comprising:

contacting a sample suspected of containing said immunological reaction partner with (1) a labeled receptor R₁ specific to said immunological reaction partner and with (2) at least one unlabeled receptor R₂ bonded to a solid phase through substance R₃, said receptor R₂ also being specific to said immunological reaction partner;

the improvement comprising carrying out a determination of the blank value by replacing R₂ by another unlabeled receptor R₂ '"'"', which does not react with the said immunological reaction partner whereby the determined blank value is used to correct for any non-specific binding.

2. Immunoassay of claim 1, wherein R₂ binds to the reaction partner to be determined.

3. Immunoassay of claim 1, wherein R₂ binds to a complex of R₁ and the reaction partner to be determined.

4. Immunoassay of claim 1, wherein R₃ is an anti-Ig antibody which does not bind to R₁.

5. Immunoassay of claim 1, wherein R₂ and R₂ '"'"' are monoclonal antibodies of the same subclass.

6. Immunoassay of claim 1, wherein R₂ and R₂ '"'"' are antibodies, each of which is bound to a hapten and R₃ is an antibody which specifically binds to said hapten.

7. A kit for the determination of a reaction partner comprising (a) a first composition containing a substance R₃ bound to a solid phase, at least one unlabelled receptor R₂ which binds to R₃ and an additional reaction partner, a labelled receptor R₂ which specifically binds to said reaction partner, and (b) a physically separated second composition used for determining a blank value containing a second portion of R₃ bound to a solid phase, a second portion of R₁, and, an unlabelled receptor R₂ which reacts with R₃ but not the further reaction partner of R₂.

8. The kit of claim 7, wherein R₂ and R₂ '"'"' are bound to a solid phase by an anti-Ig antibody.

9. The kit of claim 7, wherein R₂ and R₂ '"'"' are antibodies bound to a solid phase by an antibody specific for said hapten.