PROCESS FOR THE DEMONSTRATION AND DETERMINATION OF LOW MOLECULAR COMPOUNDS AND OF PROTEINS CAPABLE OF BINDING THESE COMPOUNDS SPECIFICALLY

• US 3,850,752 A
• Filed: 10/29/1971
• Issued: 11/26/1974
• Est. Priority Date: 11/10/1970
• Status: Expired due to Term

• Current Assignee: Antonius Hermanus Wilhelmus Maria Schuurs, Bauke Klass Van Weemen
• Original Assignee: Antonius Hermanus Wilhelmus Maria Schuurs, Bauke Klass Van Weemen
• Inventors: Schuurs, Antonius Hermanus Wilhelmus Maria, Van Weemen, Bauke Klass
• Primary Examiner(s): Tanenholtz, Alvin E.
• Application Number:
  US05/193,702
• Time in Patent Office: 1,124 Days
• Field of Search: 195/13.5R 23/253TP,23B
• US Class Current: 435/7.93
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: Y10S 435/964 including enzyme-ligand conjugate production, e.g. reducing rate of nonproductive linkage
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/815 Test for named compound or class of compounds
• CPC Class Codes: Y10S 436/817 Steroids or hormones

Abstract

The invention relates to a process for the demonstration and determination of a low molecular compound, such as a hapten, or a protein or antibody capable of binding this compound specifically, contacting a fluid containing such a compound or protein to be determined with a given quantity of the coupling product of such a low molecular compound and an enzyme, and with a given quantity of the component to be determined, which component is brought in an insoluble form, and determining the enzyme activity of the liquid or solid phase of the resulting reaction mixture.

Claims

1. PROCESS FOR THE DEMONSTRATION AND DETERMINATION OF A LOW MOLECULAR WEIGHT ORGANIC COMPOUND BY MEANS OF AN ANTIBODY AGAINST SAID LOW MOLECULAR WEIGHT COMPOUND, COMPRISING THE STEPS OF:

A. PROVIDING A GIVEN QUANTITY OF THE COUPLING PRODUCT OF SAID LOW MOLECULAR WEIGHT ORGANIC COMPOUND WITH AN ENZYME;

B. PROVIDING A CORRESPONDING GIVEN QUANTITY OF AN INSOLUBILIZED ANTIBODY AGAINST SAID LOW MOLECULAR WEIGHT ORGANIC COMPOUND;

C. CONTACTING A SAMPLE OF A FLUID CONTAING THE LOW MOLECULAR WEIGHT ORGANIC COMPOUND TO BE DETERMINED WITH SAID COMPONENTS (A) AND (B) TO FORM A REACTION MIXTURE;

AND D. DETERMINING THE ENZYME ACTIVITY OF THE LIQUID OR THE SOLID PHASE OF THE RESULTING REACTION MIXTURE, WHICH ACTIVITY IS A MEASURE OF THE QUANTITY OF LOW MOLECULAR WEIGHT ORGANIC COMPOUND TO BE DETERMINED.

2. The process of claim 1 in which said low molecular weight organic compound is a hapten.

3. The process of claim 1 in which said enzyme is an oxido-reductase.

4. PROCESS FOR THE DEMONSTRATION AND DETERMINATION OF A LOW MOLECULAR WEIGHT ORGANIC COMPOUND BY MEANS OF A PROTEIN CAPABLE OF REACTING TO BIND SAID LOW MOLECULAR WEIGHT ORGANIC COMPOUND SPECIFICALLY, COMPRISING THE STEPS OF:

A. PROVIDING A GIVEN QUANTITY OF THE COUPLING PRODUCT OF SAID LOW MOLECULAR WEIGHT ORGANIC COMPOUND WITH AN ENZUME;

B. PROVIDING A CORRESPONDING GIVEN QUANTITY OF AN INSOLUBILIZED SPECIFIC BINDING PROTEIN HAVING MORE THAN ONE VALENCY CAPABLE OF REACTING TO BIND SAID LOW MOLECULAR WEIGHT ORGANIC COMPOUND SPECIFICALLY;

C. CONTACTING A SAMPLE OF A FLUID CONTAINING THE LOW MOLECULAR WEIGHT ORGANIC COMPOUND TO BE DETERMINED WITH SAID COMPONENTS (A) AND (B) TO FORM A REACTION MIXTURE;

AND D. DETERMING THE ENZYME ACTIVITY OF THE LIQUID OR THE SOLID PHASE OF THE RESULTING REACTION MIXTURE, WHICH ACTIVITY IS A MEASURE OF THE QUANTITY OF LOW MOLECULAR WEIGHT ORGANIC COMPOUND TO BE DETERMINED.

5. The process of claim 4 in which said enzyme is an oxido-reductase.

6. PROCESS FOR THE DEMONSTRATION AND DETERMINATION OF A SPECIFIC BINDING PROTEIN HAVING TWO OR MORE BINDING SITES BY MEANS OF A LOW MOLECULAR WEIGHT ORGANIC COMPOUND CAPABLE OF REACTING TO BIND SAID PROTEIN SPECIFICALLY;

COMPRISING THE STEPS OF;

A. PROVIDING A GIVEN QUANTITY OF THE COUPLING PRODUCT OF SAID LOW MOLECULAR WEIGHT ORGANIC COMPOUND AND AN ENZYME;

B. PROVIDING A CORRESPONDING GIVEN QUANTITY OF AN INSOLUBILIZED SPECIFIC BINDING PROTEIN HAVING TWO OR MORE BINDING SITES CAPABLE OF BINDING SAID LOW MOLECULAR WEIGHT ORGANIC COMPOUND;

C. CONTACTING A SAMPLE OF A FLUID CONTAINING THE PROTEIN TO BE DETERMINED WITH SAID COMPONENTS (A) AND (B) TO FORM A REACTION MIXTURE;

AND D. DETERMINING THE ENZYME ACTIVITY OF THE LIQUID PHASE OF THE RESULTING REACTION MIXTURE, WHICH ACTIVITY IS A MEASURE OF THE QUANTITY OF THE SPECIFIC BINDING PROTEIN TO BE DETERMINED.

7. The process of claim 6 in which said protein is an antibody.

8. A diagnostic pack for the demonstration and determination of a low molecular weight organic compound by means of an antibody against said low molecular weight organic compound, comprising:

a. a given quantity of the coupling product of said low molecular weight organic compound with an enzyme;

b. a corresponding given quantity of an insolubilized antibody against said low molecular weight organic compound; and

c. a substrate for the determination of the enzyme activity of said enzyme.

9. The diagnostic pack of claim 8 in which said enzyme is an oxido-reductase.

10. A diagnostic pack for the demonstration and determination of a low molecular weight organic compound by means of a protein capable of reacting to bind said low Molecular weight organic compound specifically, comprising:

a. a given quantity of the coupling product of said low molecular weight organic compound with an enzyme;

b. a corresponding given quantity of an insolubilized specific binding protein capable of reacting to bind said low molecular weight organic compound specifically; and

c. a substrate for the determination of the enzyme activity of said enzyme.

11. The diagnostic pack of claim 10 in which said enzyme is an oxido-reductase.

12. A diagnostic pack for the demonstration and determination of a hapten by means of an antibody against said hapten, comprising:

a. a given quantity of the coupling product of said hapten with an enzyme;

b. a corresponding given quantity of an insolubilized antibody against said hapten; and

c. a substrate for the determination of the enzyme activity of said enzyme.

13. The diagnostic pack of claim 12 in which said enzyme is an oxido-reductase.