# US 5,705,400 A · Assay for adrenal autoantigen

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 5,705,400 A
- Document ID: US5705400A
- Status: Granted
- Kind: A
- Application number: 08/115,052
- Filed: 1993-09-02
- Granted: 1998-01-06
- Earliest claimed: 1992-03-07
- Assignees: RSR Limited
- Inventors: Jadwiga Maria Furmaniak-Wehr
- Web page: https://patentgrep.com/patent/US5705400A

## CPC

- 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}
- G01N 2800/04: Endocrine or metabolic disorders
- G01N 2800/24: Immunology or allergic disorders
- Y10S 436/811: Test for named disease, body condition or organ function
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

Adrenal autoantigen is isolated and characterised as a protein obtainable from the microsome fraction of the adrenal gland, having a molecular weight of about 55,000 and antigenic towards adrenal autoantibody. The protein is used in a method for detecting adrenal autoantibodies.

## Claims

### Claim 1

A method for diagnosing Addison's disease in a patient, comprising:
obtaining a sample containing antibodies from the patient; and
detecting, in the sample, the presence or absence of antibodies which specifically bind a substantially pure microsomal protein comprising an epitope for adrenal autoantibody, having an observed molecular weight of approximately 55,000 as measured by gel electrophoresis and obtained by:
homogenizing mammalian adrenal glands,
subjecting the homogenate to differential centrifugation to obtain a microsome fraction,
suspending the microsome fraction in a phosphate buffer,
centrifuging the suspension in the presence of sodium cholate to form a supernatant,
adding polyethylene glycol and further sodium cholate to the supernatant and mixing the supematant,
centrifuging the thus mixed supernatant to form a pellet,
resuspending the pellet in aqueous sodium cholate to form a suspension,
dialyzing the suspension obtained by resuspending the pellet against aqueous sodium cholate to form a solubilized microsome preparation, and
purifying the solubilized microsome preparation by column chromatography on a C.sub.8 reversed phase column equilibrated with 50 mM phosphate buffer, pH 7.0, 20% glycerol, 0.1 mM EDTA, 0.1 mM DTT, and eluted with increasing concentration of detergent to obtain a column fraction corresponding to peak VI of FIG. 1 containing the microsomal protein;
wherein presence of said antibodies in said sample is indicative of Addison's disease in the patient.