Analysis of biological fluids

• US 4,283,383 A
• Filed: 09/27/1978
• Issued: 08/11/1981
• Est. Priority Date: 05/20/1974
• Status: Expired due to Term

• Current Assignee: Technicon Instruments Corporation (Bayer AG)
• Original Assignee: Technicon Instruments Corporation (Bayer AG)
• Inventors: Masson, Pierre L., Heremans, Joseph F.
• Primary Examiner(s): Fagelson, Anna P.
• Application Number:
  US05/946,173
• Time in Patent Office: 1,049 Days
• Field of Search: 424/1, 424/1.5, 424/8, 424/11, 424/12, 424/13, 23/230 R, 23/230 B, 435/7
• US Class Current: 436/541
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• 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 436/805 Optical property
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/816 Alkaloids, amphetamines, and barbiturates
• CPC Class Codes: Y10S 436/821 involving complement factors or complement systems

Abstract

Biological fluids, such as serum, are analyzed for the presence, nature and/or amount of antibodies, antigens and antibody:antigen complexes therein using as a reagent, insolubilized rheumatoid factor or insolubilized Clq. These reagents are themselves novel. They bind to antibody:antigen complexes, but not to free antibodies or antigens, and complexes can thus be removed from mixtures thereof with other materials, such as antibodies and antigens.

Claims

1. A reagent useful in biological analysis which comprises RF or Clq covalently bonded to a solid phase substrate which is insoluble in aqueous fluids.

2. A method for quantitatively analyzing a fluid sample for an antibody -antigen complex therein, comprising:

(a) forming a mixture of the sample with a substance selected from the group consisting of RF and Clq, said substance being absorbed onto a solid phase substrate which is insoluble in aqueous fluids;

(b) allowing the substance on the substrate selectively to react with antibody -antigen complex present in the mixture thereby selectively to bind said complex to said adsorbed substance to form a second complex;

(c) assaying the second complex to determine the amount of antibody -antigen complex contained therein and thereby determining the amount of antibody -antigen complex present in the sample.

3. A method according to claim 2 wherein the solid phase is a naturally occurring material.

4. A method according to claim 3 wherein the solid phase substrate is aggregated immunoglobulins.

5. A method according to claim 2 wherein the solid phase substrate is a synthetic material.

6. A method according to claim 5 wherein the synthetic solid phase substrate is latex.

7. A method according to claim 2 wherein the substrate is in particulate form.

8. A method according to claim 2 wherein the substrate is the inner surface of a hollow tube.

9. A method according to claim 8 wherein the substrate reacts with Ab:

Ag complex by passing the fluid sample through the hollow tube.

10. A method of competitive binding assay by which the presence, in a sample, of a particular antibody, antigen or antibody -antigen complex may be detected, and the amount thereof determined, which comprises the steps of:

(a) adding to the sample to form a mixture a quantity of the specific antigen or antibody for the antibody or antigen, respectively, to be determined, or a quantity of the antibody -antigen complex to be determined, which said quantity carries an identifying label;

(b) contacting the mixture with a reagent substance selected from the group consisting of RF and Clq, said substance being adsorbed onto a solid phase substrate which is insoluble in aqueous fluids;

(c) separating the reagent substance from the mixture; and

(d) measuring by the identifying label the amount of labelled antibody, antigen, or antibody -antigen complex remaining in the mixture or separated with the reagent, thereby determining the presence and amount of antibody, antigen or antibody -antigen complex in the original sample.

11. A method for quantitatively analyzing an aqueous biological fluid sample for antibody or antigen therein, comprising:

(a) adding to the sample the appropriate antibody or antigen, respectively, to form antibody -antigen complex;

(b) forming a mixture of the sample with a substance selected from the group consisting of RF and Clq, the substance being adsorbed onto a solid phase substrate which is insoluble in aqueous fluids;

(c) allowing the substance on the substrate selectively to react with antibody -antigen complex present in the mixture thereby selectively to bind the complex to the substance; and

(d) assaying the substance to determine the amount of antibody -antigen complex bound thereto and thereby determining the amount of antibody or antigen present in the sample.

12. A method according to claim 11 wherein the solid phase is a naturally occurring material.

13. A method according to claim 12 wherein the solid phase substrate is aggregated immunoglobulins.

14. A method according to claim 11 wherein the solid phase substrate is a synthetic material.

15. A method according to claim 14 wherein the synthetic solid phase substrate is latex.

16. A method according to claim 11 wherein the substrate is in particulate form.