Method and apparatus for immunoassays

• US 4,632,901 A
• Filed: 05/11/1984
• Issued: 12/30/1986
• Est. Priority Date: 05/11/1984
• Status: Expired due to Term

• Current Assignee: Hybritech, Inc. (Danaher Corporation)
• Original Assignee: Hybritech, Inc. (Danaher Corporation)
• Inventors: Valkirs, Gunars E., Owen, Newton C., Levinson, Philip A.
• Primary Examiner(s): Marantz, Sidney
• Application Number:
  US06/609,395
• Time in Patent Office: 963 Days
• Field of Search: 435/5, 435/7, 435/287, 435/805, 435/21, 436/513, 436/527, 436/531, 436/548, 436/807, 436/808, 436/824, 436/810, 422/56-58
• US Class Current: 435/5
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/974 Aids related test
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin
• CPC Class Codes: Y10S 436/82 Hepatitis associated antigens and antibodies
• CPC Class Codes: Y10S 436/824 Immunological separation techniques

Abstract

Disclosed herein is an apparatus and process for conducting immunoassays. The apparatus comprises a first member which is a membrane or a filter to which is bound an antibody, typically a monoclonal antibody, or which is capable of extracting cells from a fluid sample. The apparatus further comprises a second member which is composed of absorbent material which acts when in contact with the first member to induce flow through the first member when a fluid sample is added to it. The apparatus is used to conduct immunoassays by applying a sample to the upper surface of the first member to bind antigen in the sample by means of antibody fixed to the first member or, in certain cases, by extracting cellular material which has antigen associated with it. Addition of the sample is followed by addition of labeled antibody against the antigen being assayed followed by a washing step to remove unbound labeled antibody. The presence of labeled antibody on the first member after washing is indicative of the presence of the antigen in the sample being assayed.

Claims

1. An apparatus for use in an immunoassay process for the detection of a target antigen in a liquid sample comprising:

(a) a first member which is a porous membrane or filter and to which is bound an antibody against the target antigen, which member has upper and lower surface, the sample being applied to the upper surface, and wherein the antibody is bound within an area smaller than the area of the member to which the sample is applied; and

(b) a second member which is a body of absorbent material having a surface over which the first member is placed and having capillaries therethrough in a direction generally transverse to the surface over which the first member is placed, which capillaries are in communication with the pores on the lower surface of the first member so as to draw liquid added to the upper surface which has permeated the first member into the capillaries of the member, the capillary communication between said first and second members having been established prior to, and maintained during, addition of liquids to the apparatus in the immunoassay process.

2. An apparatus according to claim 1 wherein the antibody is a nomoclonal antibody.

3. An apparatus according to claim 1 wherein the membrane or filter is of glass or of nylon.

4. An apparatus according to claim 1 wherein the first member is separated from the second member by another porous member.

5. An apparatus according to claim 1, 2, 3, or 4 wherein the apparatus further comprises a container for the first and second members having an opening to permit addition of assay reagents to the upper surface of the first member.

6. An apparatus according to claim 5 wherein the opening further comprises a section having sides which slope inwardly to define a funnel for direction of the added reagents onto the upper surface of the first member.

7. An apparatus according to claim 5 wherein the bottom end of the container is closed, the container being vented to allow escape of displaced air.

8. An immunoassay process for the detection of a target antigen in a liquid sample employing as an assay device an apparatus comprising:

(a) a first member which is a porous membrane or filter and to which is bound an antibody against the target antigen which member has upper and lower surfaces, the sample being applied to the upper surface, and wherein the antibody is bound within an area smaller than the area of the member to which the sample is applied; and

(b) a second member which is a body of absorbent material having a surface over which the first member is placed and having capillaries therethrough in a direction generally transverse to the surface over which the first member is placed, which capillaries are in communication with the pores on the lower surface of the first member so as to draw liquid added to the upper surface which has permeated the first member into the capillaries of the second member, the capillary communication between said first and second member having been established prior to, and maintained during, addition of liquids to the apparatus in the immunoassay process;

the process comprising adding the sample and other reagents used in the assay to the upper surface of the first member and detecting antigen bound to the antibody on the first member.

9. A process according to claim 8 wherein the sample is added first followed by addition of labeled antibody to the target antigen.

10. A process according to claim 9 wherein addition of labeled antibody is followed by washing unbound labeled antibody from the first member followed by detection of the labeled antibody, if any, bound to the first porous member.

11. A process according to claim 10 wherein the labeled antibody is labeled with an enzyme and the washing step is followed by addition to the porous member of a substrate for the enzyme which enzyme and substrate are selected to generate a color change which is either visible of determined by instrumental means.

12. A process according to claim 8, 10 or 11 wherein the antigen is selected from human choriogonadotropin, IgE, prostatic acid phosphatase, prostate specific antigens, alphafetoprotein, carcinoembryonic antigen, leutenizing hormone, creatine kinase MB, or antigens associated with hepatitis B, cytomegalovirus, herpes, and Chlamydia trachomatis.

13. A process according to claim 12 wherein the sample is urine, serum, plasma, or other liquid media.

14. A process according to claim 9, 10 or 11 wherein the bound and labeled antibodies are monoclonal antibodies which bind, respectively, to non-interfering epitopes on the antigen.

15. A process according to claim 14 wherein the labeled antibody is labeled with an enzyme.

16. A process according to claim 15 wherein a solution of enzyme substrate is added after addition of the labeled antibody.