Method for carrying out immunodiagnostic tests

• US 5,420,047 A
• Filed: 11/12/1993
• Issued: 05/30/1995
• Est. Priority Date: 11/13/1992
• Status: Expired due to Fees

• Current Assignee: Bayer AG
• Original Assignee: Bayer AG
• Inventors: Brandt, Heinz-Dieter, Stocker, Ronald, Hildenbrand, Karlheinz, Dhein, Rolf
• Primary Examiner(s): Bidwell, Carol E.
• Application Number:
  US08/149,247
• Time in Patent Office: 564 Days
• Field of Search: 435/7.9, 435/7.92, 435/7.94, 435/970, 435/805, 435/25, 435/28, 435/4, 436/518, 436/528, 436/531, 436/810, 436/532, 422/56, 422/57, 422/60.1, 422/69
• US Class Current: 435/7.9
• CPC Class Codes: C12N 11/082 obtained by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: C12N 11/087 Acrylic polymers
• CPC Class Codes: C12N 11/089 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: C12N 11/093 Polyurethanes
• CPC Class Codes: C12N 11/12 Cellulose or derivatives thereof
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 33/545 Synthetic resin
• 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

Abstract

Immunodiagnostic tests in which enzymes or antibodies are bound to porous carrier materials can be carried out by employing as porous carrier materials macroporous polymer membranes which are prepared by

(a) dispersing an insoluble filler in a solution which contains at least two incompatible polymers in amounts which lead to phase separation in the solution, resulting in a homogeneous casting solution, and

(b) applying this solution to a support and carrying out a precipitation coagulation.

Claims

1. In an immunoassay comprising contacting a macroporous membrane to which an enzyme or antibody is absorbed with a liquid putatively containing an antigen and detecting the presence of the antigen, the improvement comprising utilizing as the macroporous membrane a macroporous filler-containing membrane where the membrane is prepared by (1) forming an unstable mixture of at least two polymers which are immisible in solution,

(2) dispersing insoluble fillers into the unstable mixture of such immisible polymers thereby converting the mixture into a stable homogeneous mixture, and

(3) applying such stable mixture to a substrate as a casting solution by precipitation coagulation.

2. The method of claim 10, wherein the insoluble fillers comprise from 5 to 500% by weight based on the total weight of the polymers in the membrane.

3. The method of claim 1, wherein the fillers are highly dispersed fillers and are employed in amounts of 5 to 50% by weight.

4. The method of claim 1, wherein the fillers have lower specific surface areas and are employed in amounts of 35 to 500% by weight.

5. The method of claim 1, wherein the polymers are selected from the group consisting of combinations of

a) cellulose esters/polyvinyl esters,

b) polyurethane/polyacrylates or acrylic copolymers,

c) polycarbonate copolymers/polyurethane,

d) polyvinyl derivatives/polysulphones,

e) polyamides or polyimides/polystyrene or styrene copolymers,

f) poly-para-dimethylphenylene oxide/polyvinylidene fluoride,

g) polysulphone/polyacrylonitrile, and

h) poly(ethersulphone)/anionically modified polyacrylonitrile.

6. The method of claim 1 wherein the macroporous polymer membrane contains an integrated chromogen.

7. The method of claim 1, wherein the macroporous membrane has a surface modified by free amino groups.

9. The method of claim 1, wherein the insoluble fillers are selected from the group consisting of talc, titanium dioxide, barium sulphate, silicon dioxide, microcrystalline cellulose, zeolite, bentonite, highly dispersed silica and quartz.

10. The immunoassay of claim 1 wherein the macroporous filler-containing membrane contains free amino groups.

11. The immunoassay of claim 1 wherein the filler in the macroporous filler-containing membrane is an amino-modified filler.

12. The immunoassay of claim 1 wherein the enzyme or antibody is attached to the membrane through a covalent linkage.

13. The method of claim 1, wherein the forming step comprises forming an unstable mixture of three immiscible polymers.

8. The method of claim 8, wherein bisulphite adducts of isocyanates are employed for the covalent binding of the enzymes or antibodies to the free amino groups on the surface of said macroporous membrane.