Detection of organic compounds through regulation of antibody-catalyzed reactions

• US 5,736,343 A
• Filed: 08/16/1996
• Issued: 04/07/1998
• Est. Priority Date: 08/18/1995
• Status: Expired due to Fees

• Current Assignee: Donald Landry
• Original Assignee: Donald Landry
• Inventors: Landry, Donald
• Primary Examiner(s): Patterson, Jr., Charles L.
• Application Number:
  US08/698,812
• Time in Patent Office: 599 Days
• Field of Search: 435/7.6, 435/188.5
• US Class Current: 435/7.6
• CPC Class Codes: G01N 2333/901 Antibodies with enzymatic activity; e.g. abzymes
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/577 involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens; G01N33/53 - G01N33/576 take precedence
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92

Abstract

The present invention provides a method for determining the presence of a target organic molecule in a sample which comprises either (A): first, adding a substrate molecule to the sample, the substrate molecule being a conjugate of the target organic molecule and a reporter molecule; second, adding a catalytic monoclonal antibody to the sample which (1) recognizes and binds the target organic molecule when the target organic molecule is present in the sample and (2) recognizes and binds the substrate molecule when the target organic molecule is not present in the sample; and third detecting a change in the sample signifying the absence of the target organic molecule, the change being the product of an antibody-catalyzed reaction and thereby determining the presence of an organic molecule in the sample; or (B): first adding a substrate molecule to the sample, the substrate molecule being a conjugate of a molecule complementary to the target molecule and a reporter molecule; second adding a catalytic monoclonal antibody to the sample which (1) recognizes and binds the target organic molecule when the target molecule is present in the sample and (2) recognizes and binds the substrate molecule only if the target molecule is present in the sample; and third detecting a change in the sample signifying the presence of the target molecule, the change being the product of an antibody-catalyzed reaction and thereby determining the presence of the target organic molecule in the sample.

Claims

1. A method for determining the presence or absence of a target organic molecule in a sample which comprises:

a. adding a substrate molecule to the sample,

b. adding a catalytic monoclonal antibody to the sample which

(i) recognizes and binds the target organic molecule when the target organic molecule is present in the sample, or

(ii) recognizes, binds, and transforms the substrate molecule when the target organic molecule is not present in the sample; and

c. detecting a change in the sample signifying the absence of the target organic molecule, the change being the production of a reporter molecule by an antibody-catalyzed reaction of the substrate molecule and thereby determining the presence or absence of the target organic molecule in the sample.

3. The method of claim 1 or 2, wherein the target organic molecule is phenylalanine.

4. The method of claim 1 or 2, wherein the sample is derived from a cell sample.

5. The method of claim 1 or 2, wherein the sample is derived from a tissue sample.

6. The method of claim 1 or 2, wherein the sample is a biological fluid sample.

7. The method of claim 1 or 2, wherein the sample is from a human subject.

8. The method of claim 1 or 2, wherein the sample is from an animal subject.

9. The method of claim 8 wherein the animal subject is a dog, a horse, a cat, a cow or a pig.

10. The method of claim 1 or 2, wherein the reporter molecule is a detectable marker.

11. A method of claim 10, wherein the detectable marker contains a radioactive label.

12. A method of claim 11, wherein the radioactive label is ¹²⁵ I.

13. A method of claim 10, wherein the detectable marker is a colorometric marker.

14. A method of claim 10, wherein the detectable marker is a fluorometric marker.

15. A method of claim 10, wherein the detectable marker is a chemiluminescent or bioluminescent marker.

16. A method of claim 10, wherein the detectable marker is the product of an enzymatic reaction.

17. The method of claim 1 or 2, wherein the catalytic monoclonal antibody recognizes and binds phenylpyruvate.

18. The method of claim 1 or 2, wherein the change is a visual change.

19. The method of claim 18, wherein the visual change is a change in the color of the sample.

20. A method of claim 1 or 2, wherein the catalytic monoclonal antibody is bound to a matrix.

21. A method of claim 20, wherein the matrix is agarose or sepharose.

22. A method of claim 1 or 2, wherein the catalytic monoclonal antibody is bound to a tube.

23. A method of claim 1 or 2, wherein the catalytic monoclonal antibody is bound to a bead.

24. A method for diagnosing in a subject a disease condition associated with the presence of an organic molecule which comprises determining in a sample from the subject the presence of the organic molecule in accordance with the method of claim 1 or 2, the presence of the organic molecule indicating the presence of the disease condition.

25. A method for monitoring the course of a disease condition in a subject which comprises determining in a first sample from the subject the presence of an organic molecule according to the method of claim 1 or 2 and comparing the amount so determined with the amount present in a second sample from the subject, such samples being taken at different points in time, a difference in the amounts determined being indicative of the course of the disease condition.

2. A method for determining the presence of a target organic molecule in a sample which comprises:

a. adding a catalytic monoclonal antibody to the sample which

(i) recognizes and binds the target organic molecule when the target organic molecule is present in the sample and

(ii) recognizes, binds and transforms a substrate molecule when the target organic molecule is present in the sample;

b. adding a substrate which is the conjugate of a reporter molecule and a molecule which complements the target organic molecule through non-covalent bonding interactions;

c. detecting a change in the sample signifying the presence of the target organic molecule, the change being the production of the reporter molecule by an antibody-catalyzed reaction of substrate molecule and thereby determining the presence of target organic molecule in the sample.