Immunoassay for determination of immune complexes with polymer-coated plastic base

• US 4,450,231 A
• Filed: 03/31/1982
• Issued: 05/22/1984
• Est. Priority Date: 03/31/1982
• Status: Expired due to Term

• Current Assignee: REAADS Medical Products, Inc. (Sebia SA)
• Original Assignee: Biostar Medical Products Incorporated
• Inventors: Ozkan, Adil N.
• Primary Examiner(s): Marantz, Sidney
• Application Number:
  US06/363,967
• Time in Patent Office: 783 Days
• Field of Search: 435/7, 435/293, 436/506, 436/538, 436/539, 436/821, 436/507, 436/529, 436/531, 436/809
• US Class Current: 435/7.92
• CPC Class Codes: G01N 33/544 the carrier being organic
• CPC Class Codes: G01N 33/564 for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
• CPC Class Codes: Y10S 435/967 Standards, controls, materials, e.g. validation studies, buffer systems
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays

Abstract

An immunoassay of a specimen of a serum or the like to determine immune complexes, includes producing on a plastic base a layer of a non-proteinaceous, non-ionic polymer which will adhere to the plastic base and has the capability of absorbing immune complexes of the specimen, placing a specimen on the layer and treating the layer to produce an indication of the amount of immune complexes. The polymer may be polyethylene glycol, dextran, polyvinyl chloride, a polymeric polyol or an adduct of polyethylene glycol. Washing with conventional solutions, addition of an antihuman IgG coupled with an enzyme and addition of a substrate reactive therewith, are similar to the ELISA test, with color measurement as by spectrophotometer. Or, the addition of anti IgG-I¹²⁵ and measurement by a scintillation counter may be used. The ethylene glycol may range in molecular weight from 2,000 to 20,000, although 6,000 to 8,000 is preferred. A product for use in such an assay is a plate having wells or a test tube formed of plastic, polystyrene and polyvinyl chloride being preferred, with a layer of such non-proteinaceous, non-ionic layer on the plate wells or the cavity of the test tube. This product, if it is not to be used immediately, should be provided with a layer or film protecting the polymer layer from the air.

Claims

1. In a method of performing an immunoassay of a fluid specimen to determine immune complexes therein, the steps comprising:

treating a solid phase, plastic base having means for receiving the fluid specimen to be assayed, to produce a layer of a non-proteinaceous non-ionic polymer on said means, said polymer adhering to said plastic base and having the capability of absorbing immune complexes which may be present in such specimen;

placing the fluid specimen to be assayed on said receiving means;

permitting said polymer layer to absorb immune complexes present in said specimen; and

treating said layer in a manner to produce an indication of the amount of immune complexes in said layer.

2. In a method as defined in claim 1, wherein:

said polymer is selected from the group consisting of polyethylene glycol, dextran and polyvinyl chloride.

3. In a method as defined in claim 1, wherein:

said non-ionic polymer is polyethylene glycol having a molecular weight from about 2000 to about 20,000.

4. In a method as defined in claim 1, wherein:

said polymer is a polymeric polyol.

5. In a method as defined in claim 1, wherein:

said indication treatment produces a change in color, generally proportional to the amount of immune complexes absorbed by said layer; and

measuring the color so produced by visual or spectrophotometric means.

6. In a method as defined in claim 5, which includes:

washing said base a plurality of times with a buffer solution following production of said polymer layer thereon;

washing said plate, after absorption of immune complexes of said specimen by said polymer layer, a plurality of times with a buffer solution;

adding an anti-human IgG coupled with an enzyme to said layer and permitting said layer to stand for a predetermined time;

washing said layer a plurality of times with a buffer solution after said layer is permitted to stand for said predetermined time;

adding a substrate reactive with said anti-human IgG coupled with an enzyme to produce a color change proportional to the amount of said anti-human IgG coupled with an enzyme which has reacted with immune complexes present in said layer; and

measuring the color change in said layer by means of a spectrophotometer.

7. In a method as defined in claim 1, wherein:

said indication treatment involves a radioactive agent which produces a radiation intensity generally proportional to the amount of immune complex absorbed by said layer; and

measuring the amount of radiation present.

8. In a method as defined in claim 1, which includes:

treating a second fluid specimen, essentially identical to said first specimen, with a virus or bacteria of a suspected condition; and

performing an immunoassay on said second specimen simultaneously with said first specimen.

9. In a method of performing an immunoassay of a fluid specimen to detect immune complexes therein, the steps comprising:

placing the fluid specimen on a receiving means of a solid phase, plastic base having at least one receiving means, with said receiving means having a layer of a non-proteinaceous, non-ionic polymer adhering thereto and having the capability of absorbing immune complexes from such specimens;

placing the fluid specimen to be assayed on said receiving means;

permitting said polymer layer to absorb such immune complexes; and

treating said layer in a manner to provide an indication of the amount of immune complexes in said layer.

10. In a method as defined in claim 9, wherein:

said polymer is selected from the group consisting of polyethylene glycol, dextran and polyvinyl chloride.

12. In a method as defined in claim 9, wherein:

said polymer is a polymeric polyol.

13. In a method as defined in claim 9, wherein:

said indication treatment produces a change in color, generally proportional to the amount of immune complexes absorbed by said layer; and

measuring the color so produced.

14. In a method as defined in claim 13, which includes;

washing said base a plurality of times with a buffer solution following production of said polymer layer thereon;

washing said plate, after absorption of immune complexes of said specimen by said polymer layer, a plurality of times with a buffer solution;

adding an anti-human IgG coupled with an enzyme to said layer and permitting said layer to stand for a predetermined time;

washing said layer a plurality of times with a buffer solution after said layer is permitted to stand for said predetermined time;

adding a substrate reactive with said anti-human IgG coupled with an enzyme to produce a color change proportional to the amount of said anti-human IgG coupled with an enzyme which has reacted with immune complexes present in said layer; and

measuring the color change in said layer by means of a spectrophotometer.

15. In a method as defined in claim 9, wherein:

said indication treatment involves a radioactive agent which produces a radiation intensity generally proportional to the amount of immune complex absorbed by said layer; and

measuring the amount of radiation present.

16. In a method as defined in claim 9, which includes:

treating a second fluid specimen, essentially identical to said first specimen, with a virus or bacteria of a suspected condition; and

performing an immunoassay on said second specimen simultaneously with said first specimen.

11. In a method as defined in clim 9, wherein:

said non-ionic polymer is polyethylene glycol having a molecular weight from about 2000 to about 20,000.