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Methods of detecting autoantibodies for diagnosing and characterizing disorders

A technology for autoantibodies and disorders, applied in the fields of disease diagnosis, chemical method analysis, measurement devices, etc., can solve the problem of not being able to truly detect the early stage of the target disorder

Inactive Publication Date: 2010-02-24
UNIV OF LOUISVILLE RES FOUND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Therefore, current antigen-based assays cannot truly detect early stages of target disorders
To significantly improve diagnostic assays for the disorder of interest, there is an unmet need for a new method for detecting the disorder that is more sensitive to the presence of disease markers early in its establishment

Method used

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  • Methods of detecting autoantibodies for diagnosing and characterizing disorders
  • Methods of detecting autoantibodies for diagnosing and characterizing disorders
  • Methods of detecting autoantibodies for diagnosing and characterizing disorders

Examples

Experimental program
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Effect test

Embodiment 1

[0125] Reactive Antibodies Linked to Cancer

[0126] Autoreactive antibodies have been shown to be prevalent in all cancer patients tested. The presence of tumor-reactive autoantibodies was analyzed in ovarian cancer patients (n=28) and tumor-free age-matched female volunteers (n=32). The presence of IgG in ovarian cancer patient sera and their reactivity with ovarian tumor-derived cellular antigens were quantified by ELISA.

[0127] Tumor-derived cell antigens from UL-1 ovarian tumor cells in logarithmic growth phase were diluted 1:25 into coupling buffer (pH 8.3) consisting of 100 mM sodium carbonate and 0.5 M NaCl. Aliquots were added to wells of a 96-well Immulon4 microtiter plate and incubated overnight at 37°C. The plates were blocked with 5% non-fat dry milk and then incubated overnight at 4°C with 200 μl of serum diluted 1 / 200 from ovarian cancer patients and age-matched normal controls. The plate was washed and incubated with peroxidase-conjugated human IgG antibod...

Embodiment 2

[0131] Specific antigen target recognition by tumor-reactive IgG

[0132] Since the reactivity of autoantibodies from ovarian cancer patient sera to ovarian tumor antigens can exhibit quantitative differences based on the stage of the disease, qualitative differences were also investigated using Western blot and densitometry. Using cellular protein antigens from three ovarian tumor cell lines (UL-1, UL-3, and UL-6) in logarithmic growth phase, the activity of components reactive with the humoral immune response of ovarian cancer patients was assessed by Western blotting. exist( figure 2 , representative blot). Western blotting was performed using patient serum (1:100 dilution) as the primary antibody source. The binding of the patient-derived antibody to the tumor cell-derived antigen was visualized using ECL, first using a peroxidase-conjugated human IgG antibody. Immunoreactivity was quantified by densitometry. The antigens recognized and differences in the strength of ...

Embodiment 3

[0138] Marker Identification for Treatment Response

[0139] Since differences in reactivity patterns occurred in patients with advanced disease, the correlation between this difference and chemoresistance was examined. Separation of cellular components by 2-dimensional electrophoresis followed by western blotting allowed some of these differences to be assessed. Identification of a cluster of 3 points associated with cisplatin resistance was identified using sera from patients who did not initially respond to cisplatin therapy and patients who presented an initial response and remained disease-free for >12 months ( image 3 ). Utilization of these cellular antigens as part of the diagnostic arrays disclosed herein allows early identification of resistant tumors.

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Abstract

Methods for detecting and / or quantitating levels of autoantibodies in subjects are provided. Methods for diagnosing and / or characterizing a disorder associated with autoantibody production are furtherprovided. In some embodiments, the disorder diagnosed and / or characterized can be a cancer or an infertility disorder.

Description

[0001] related application [0002] The subject matter disclosed in this application claims priority based on US Provisional Patent Application Serial No. 60 / 897,641, filed January 26, 2007; the disclosure of which provisional application is incorporated herein by reference in its entirety. [0003] Government interests [0004] The subject matter disclosed in this application was made in part with United States Government support under Grant No. CA104651 awarded by the National Cancer Institute. Accordingly, the United States Government has certain rights in the subject matter disclosed in this application. technical field [0005] The subject matter disclosed herein relates to the diagnosis and characterization of disorders in subjects by detection of autoantibodies. Specifically, the subject matter disclosed herein relates to the use of antigens (including, in some embodiments, antigens isolated from exosomes produced by cells) to detect autoantibodies in a subject for th...

Claims

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Application Information

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IPC IPC(8): G01N31/00G01N33/543
CPCG01N2800/367G01N33/6854G01N33/574
Inventor D·D·泰勒C·格塞欧-泰勒
Owner UNIV OF LOUISVILLE RES FOUND INC
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