Enzyme immunoassay with two-zoned device having bound antigens

• US 4,446,232 A
• Filed: 10/13/1981
• Issued: 05/01/1984
• Est. Priority Date: 10/13/1981
• Status: Expired due to Term

• Current Assignee: Lance A. Liotta
• Original Assignee: Lance A. Liotta
• Inventors: Liotta, Lance A.
• Primary Examiner(s): Marantz, Sidney
• Application Number:
  US06/310,801
• Time in Patent Office: 931 Days
• Field of Search: 435/4, 435/7, 435/188, 435/810, 435/805, 422/55, 422/56, 422/57, 422/58, 422/61, 436/514, 436/515, 436/518, 436/528, 436/529, 436/530, 436/531, 436/532, 436/535, 436/63, 436/516
• US Class Current: 435/7.93
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 33/54387 Immunochromatographic test strips
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin
• CPC Class Codes: Y10S 436/82 Hepatitis associated antigens and antibodies

Abstract

A device for determining the presence of antigens which comprises a first zone containing antigens and enzyme-linked antibodies which are capable of immunologically reacting with said antigens, said antibodies being positioned in said first zone such that they will be removed from said first zone when reacted with antigens passing through said first zone but not removed from said first zone in the absence of such antigens, and a second zone containing material capable of reacting with said enzyme-linked antibodies to produce a color forming reaction which indicates the presence of said antibodies.

Claims

1. A device for determining the presence of antigens which comprises:

a matrix;

a first zone in said matrix containing (a) bound and immobilized antigens and (b) enzyme-linked antibodies which are capable of immunologically reacting with said antigens, said antibodies being positioned in said first zone such that they will be removed from said first zone when reacted with antigens passing through said first zone but not removed from said first zone in the absence of such antigens; and

a second zone removed from said first zone containing material capable of reacting with said enzyme-linked antibodies to produce a color forming reaction which indicates the presence of said antibodies.

2. The device of claim 1 wherein said first and second zones are in a juxtapositioned relationship.

3. The device of claim 1 wherein said first zone contains enzyme-linked antibodies which are combined with bound and immobilized specific antigens.

4. The device of claim 1 wherein said first zone consists of at least two individual layers, with one of said layers containing enzyme-linked antibodies and another layer containing specific antigen bound and immobilized in said layer.

5. The device of claim 1 wherein said first zone is fabricated from nitrocellulose.

6. The device of claim 1 wherein said second zone is fabricated from nitrocellulose.

7. The device of claim 1 wherein said first zone is fabricated from a fibrous material.

8. The device of claim 1 wherein said second zone is fabricated from a fibrous material.

9. The device of claim 1 wherein said matrix is fabricated from a fibrous material.

10. The device of claim 9 wherein said fibrous material is diazobenzyloxymethyl paper.

11. The device of claim 1 wherein said first zone is fabricated from a porous polymer gel.

12. The device of claim 11 wherein said porous gel is at least one material selected from the group consisting of polyacrylamide, agarose and collagen gels.

13. A method for determining the presence of antigens in a biological fluid which comprises:

bringing a fluid which is to be tested for the presence of antigens into contact with a device having a matrix including a first zone containing (a) bound and immobilized antigens and (b) enzyme-linked antibodies which are capable of immunologically reacting with said antigens, said antibodies being positioned in said first zone such that they will be removed from said first zone but not removed from said first zone in the absence of such antigens, and a second zone removed from said first zone containing material capable of reacting with said enzyme-linked antibodies to produce a color forming reaction which indicates the presence of said antibodies;

allowing said fluid to permeate said device; and

observing the presence or absence of any color change in said second zone to thereby determine the presence or absence of the tested for antigen in said fluid.

14. The method of claim 13 wherein said first and second zones are in a juxtapositioned relationship.

15. The method of claim 13 wherein said first zone consists of at least two individual layers, with one of said layers containing enzyme-linked antibodies and another layer containing specific antigen bound and immobilized in said layer.

16. The method of claim 13 wherein said first zone consists of at least two individual layers, with one of said layers containing enzyme-linked antibodies and another layer containing specific antigen bound and immobilized in said layer.

17. The method of claim 13 wherein said first zone is fabricated from nitrocellulose.

18. The method of claim 13 wherein said second zone is fabricated from nitrocellulose.

19. The method of claim 13 wherein the antigen being determined is HCG and wherein the antigen in said device is human chorionic gonadatrophin.

20. The method of claim 13 wherein the antigen being determined is hepatitis antigen and wherein the antigen in said device is hepatitis antigen.

21. The method of claim 13 wherein the antigen being determined is rubella virus protein and wherein the antigen in said device is a rubella virus protein.

22. A device for determining the presence of antibodies which comprises:

a matrix;

a first zone is said matrix containing enzyme-linked antigens and bound and immobilized antibodies which are capable of immunologically reacting with said enzyme-linked antigens, said enzyme-linked antigens being positioned in said first zone such that they will be removed from said first zone but not removed from said first zone in the absence of such antibodies; and

a second zone removed from said first zone containing material capable of reacting with said enzyme-linked antigens to produce a color forming reaction which indicates the presence of said antigens.

23. A method for determining the presence of antibodies in a biological fluid which comprises:

bringing a fluid which is to be tested for the presence of antibodies into contact with a device having a matrix including a first zone containing enzyme-linked antigens and bound and immobilized antibodies which are capable of immunologically reacting with said enzyme-linked antigens, said enzyme-linked antigens being positioned in said first zone such that they will be removed from said first zone when reacted with antibodies passing through said first zone but not removed from said first zone in the absence of such antibodies, and a second zone removed from said first zone containing material capable of reacting with said enzyme-linked antigens to produce a color forming reaction which indicates the presence of said antigens;

allowing said fluid to permeate said device; and

,

observing the presence of absence of any color change in said second zone to thereby determine the presence or absence of the tested for antibodies in said fluid.