Antibody assay

A technology of antibodies and autoantibodies, which is applied in the direction of measuring devices, biological testing, material inspection products, etc.

Active Publication Date: 2018-06-01
ONCIMMUNE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its poor performance highlights a major gap in improved early detection / screening tests for liver cancer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0218] Example 1 - General protocol for measuring autoantibodies to tumor-associated proteins

[0219] Samples of tumor marker antigens can be prepared by recombinant expression following methods similar to those described in WO 99 / 58978, the contents of which are incorporated herein by reference. Briefly, cDNA encoding the marker antigen of interest was cloned into the pET21 vector (Invitrogen) modified to encode a biotin tag and a 6x histidine tag to facilitate purification of the expressed protein. The resulting clones were grown in BL21(DE3) E. coli and the bacteria were subsequently lysed. Expressed antigen was recovered by nickel chelate affinity column (HiTrap, commercially available from GE Healthcare) following the manufacturer's protocol. Purity, specificity and yield of expressed protein were assessed by SDS-PAGE, Western blot and protein assay prior to storage.

[0220] Negative control protein VOL was generated by transforming BL21(DE3) E. coli with an empty pET...

Embodiment 2

[0268] Example 2 - Detection of autoantibodies in hepatocellular carcinoma (HCC) by HTPA

[0269] The following data were obtained from a pilot study evaluating the sensitivity and specificity of a panel of autoantibody assays in HCC detection using the HTPA format. The clinical and demographic status of the subjects included in the study are given in Tables 1 to 4.

[0270] Table 1 - Demographics of patients included in the study described in Example 2

[0271]

[0272]

[0273] Table 2 - Size of primary tumors present in HCC patients

[0274]

number of patients available

average value

min-max

Primary tumor size (cm)

88

5.9

0.4-19

[0275] Table 3 - Tumor staging of HCC patients using TNM staging

[0276] TNM staging

number

1

35

2

21

3

21

4

2

N / A

20

[0277] TNM = TNM classification of the malignancy staging system; N / A = not available

[0278] Table 4 - Liver ...

Embodiment 3

[0302] Example 3 - Additional measurement of AFP in combination with autoantibodies measured by HTPA

[0303] Circulating alpha-fetoprotein (AFP) was measured in serum samples using a commercially available ELISA (Aviva Systems Biology) for the sample set described in Example 2. A common cut-off of 200 ng / ml was used to assess positivity. Table 12 shows the results of adding AFP to Group 1 and Group 2 shown in Example 2. From these results, it is clear that the performance of AFP combined with AAb group is greater than that of AFP or AAb group alone.

[0304] Table 12 - Performance of AFP alone and when added to the panel described in Example 2

[0305]

[0306]

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Abstract

The present invention relates to a method of detecting liver cancer in a mammalian subject by detecting an antibody in a test sample comprising a bodily fluid from the mammalian subject, wherein the antibody is an autoantibody immunologically specific for a tumour marker protein selected from the group consisting of MMP9, AIF1, EpCAM and CDKN1B, which method comprises contacting the test sample with a tumour marker antigen selected from the group consisting of MMP9, AIF1, EpCAM and CDKN1B and determining the presence or absence of complexes of the tumour marker antigen bound to autoantibodiespresent in the test sample where the presence of said complexes is indicative of the presence of liver cancer. Also included within the invention are corresponding methods of diagnosing and treating liver cancer in a mammalian subject, corresponding methods of predicting response to an anti-liver cancer treatment, a corresponding method of detecting an antibody in a test sample comprising a bodilyfluid from a mammalian subject and kits suitable for performing methods of the invention.

Description

technical field [0001] The present invention relates generally to the field of antibody detection, and in particular to assays for the detection of liver cancer-associated autoantibodies in samples comprising bodily fluids of patients. Background technique [0002] Many diagnostic, prognostic and / or monitoring assays rely on the detection of biomarkers of a particular disease state or disease susceptibility. Such biomarkers are typically proteins or polypeptides that are characteristic of a particular disease or associated with disease susceptibility, and are often used to detect cancer, including liver cancer. [0003] Liver cancer and especially hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide and it is also the second most common cause of cancer death. High mortality is caused by late diagnosis (often after metastasis) and pre-existing liver disease. Late diagnosis is due to lack of early symptoms and suboptimal imaging techniques used for diagno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/57438G01N33/57484G01N33/6893G01N2800/50G01N2800/52G01N33/564
Inventor 贾里德·艾伦伊莎贝尔·麦克唐纳安德烈亚·默里克里斯托弗·韦尔拜里
Owner ONCIMMUNE
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