Membrane-supported immunoassays

• US 4,874,691 A
• Filed: 10/16/1987
• Issued: 10/17/1989
• Est. Priority Date: 10/16/1987
• Status: Expired due to Fees

• Current Assignee: Quadra Logic Technologies Incorporated
• Original Assignee: Quadra Logic Technologies Incorporated
• Inventors: Chandler, Howard
• Primary Examiner(s): Nucker, Christine M.
• Application Number:
  US07/109,436
• Time in Patent Office: 732 Days
• Field of Search: 436/518, 436/533, 436/534, 436/807, 436/535, 435/7, 422/61, 422/58, 422/101, 422/64
• US Class Current: 435/7.92
• CPC Class Codes: G01N 2035/1055 for immobilising reagents, e.g. dried reagents
• CPC Class Codes: G01N 33/5436 with ligand physically entrapped within the solid phase liposomes G01N33/5432; immunological test elements G01N33/54386
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/76 Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures

Abstract

Improved, economical devices for immunoassays employ a reusable syringe or vacuum manifold to withdraw samples through a membrane containing an affinity partner for analyte. The devices can also be adapted to direct blood sampling and to automated assays.

Claims

1. A method to conduct an assay for an analyte in a sample, wherein the analyte binds to an affinity partner, which method comprises:

drawing a volume of said sample by means of a pressure gradient through a membrane in which the affinity partner for the analyte is contained into a receiving chamber on which said membrane is mounted,

wherein said pressure gradient is obtained by creating a partial vacuum in said receiving chamber, and

detecting the analyte bound in the membrane.

2. The method of claim 1 wherein the receiving chamber is the body of a syringe.

3. The method of claim 1 wherein the receiving chamber is a vacuum manifold.

4. The method of claim 1 wherein the affinity partner for the analyte is a specific affinity partner.

5. The method of claim 1 wherein the affinity partner for the analyte is an immunoglobulin or derivative thereof.

6. The method of claim 1 wherein the affinity partner for the analyte is an antigen or hapten.

7. The method of claim 1 wherein the bound analyte is detected by reaction with a labeled immunoglobulin or derivative thereof.

8. The method of claim 7 wherein the immunoglobulin or derivative thereof is an antibody specific for the analyte.

9. The method of claim 8 wherein the labeled antibody is labeled with an enzyme.

10. The method of claim 2 wherein the sample is blood or internal body fluids drawn directly past the membrane.

11. A method to conduct an assay for analyte in two or more samples simultaneously wherein the analyte binds to an affinity partner, which method comprises:

placing a volume of each sample in contact with one of a multiplicity of membranes in each of which the affinity partner for the analyte is contained,

wherein each of said multiplicity of said membranes is mounted onto a common receiving chamber, and

in a single operation withdrawing each sample by means of a pressure gradient applied as a partial vacuum in said receiving chamber through the membrane into said receiving chamber, and

detecting the analyte bound in each membrane from each sample.

12. The method of claim 11 wherein each sample is placed in an outer chamber affixed to the corresponding membrane and wherein said pressure gradient is obtained by creating a partial vacuum in said receiving chamber.

13. An apparatus for the detection of analyte in a sample wherein the detection comprises reaction of said analyte with an affinity partner, which apparatus comprises:

a supported membrane containing said affinity partner, wherein said membrane is supported at the upper portion of a housing wherein said housing further includes means to attach said membrane to an orifice of a receiving chamber in which a partial vacuum can be created.

14. The apparatus of claim 13 which further includes the body of a syringe attached to the housing.

15. The apparatus of claim 13 wherein the affinity partner is a specific affinity partner.

16. The apparatus of claim 13 wherein the affinity partner is an immunoglobulin or derivative thereof.

17. The apparatus of claim 13 wherein the affinity partner is an antigen or hapten.

18. The apparatus of claim 16 wherein the immunoglobulin or derivative thereof is an antibody specific for the analyte.

19. The apparatus of claim 13 wherein the housing supports the membrane in a horizontal position.

20. The apparatus of claim 13 wherein said housing engages said membrane by means of a deformable lip.

21. The apparatus of claim 13 wherein the membrane is engaged at the top face of the housing.

22. The apparatus of claim 13 which further includes an outer chamber disposed at the top of the housing capable of accommodating the sample.

23. The apparatus of claim 13 wherein said apparatus comprises an upper portion and a lower portion,

wherein said lower portion is a luer socket contiguous with a lip portion and

wherein said upper portion is a cylindrical chamber having at one end a flange of dimension to engage said lip of the lower portion and

wherein said membrane is supported between the flange of the upper portion and lip of the lower portion.

24. The apparatus of claim 13 wherein said apparatus has an upper portion and a lower portion,

wherein said lower portion is a luer socket having at one end an extended rim on which said membrane is supported,

said upper portion fitting annularly at the exterior surface of the lower portion and having at the top thereof a lip extension which engages the membrane supported on the lower portion.

25. The apparatus of claim 22 wherein said apparatus has a top portion and bottom portion,

wherein said top portion is a cylindrical chamber containing at its base said membrane,

said cylindrical chamber having an engaging means adjacent said membrane for attachment of said top portion to said bottom portion,

wherein said bottom portion contains an engaging means for the top portion and a means for attachment of the apparatus to a vacuum manifold.

29. A kit suitable for conducting measurement of an analyte in a biological sample which reagent kit comprises:

the supported membrane unit of claim 13, reagents in separate bottles, and instructions for conducting the assay.

30. The kit of claim 29 which further includes a syringe

31. The kit of claim 29 which further includes a vacuum manifold.

32. The kit of claim 30 wherein the syringe is reusable.

26. A method to conduct an assay for analyte in a sample, wherein the analyte binds to a first affinity partner and wherein said binding is detected by reaction with a labeled second affinity partner, wherein at least one of said first and second affinity partners is specific for analyte, which method comprises:

drawing a volume of said sample by means of a pressure gradient through a porous support containing said labeled second affinity partner and through a membrane in which the first affinity partner is contained into a receiving chamber, wherein said pressure gradient is obtained by creating a partial vacuum in said receiving chamber.

27. An apparatus for conducting an assay for analyte in a sample which comprises:

a needle;

attached to a housing in which is supported a membrane containing an affinity partner for the analyte, which membrane is connected to a receiving chamber which includes a means of applying a pressure gradient through the needle and past the membrane into the chamber.

28. An apparatus for conducting an assay for an analyte in a sample which comprises a means for applying a pressure gradient past a membrane in which a first affinity partner for the analyte is contained, into a receiving chamber and having upstream of said membrane a porous support containing a second affinity partner conjugated to label, wherein at least one of said first and second affinity partners is specific for analyte.