Molecular marker for early indentification of pleural mesothelioma patients, and expression analysis method for same

A technology for skin tumors and patients, applied in the field of mesothelioma inspection, can solve problems such as the unknown relationship between periostin and mesothelioma

Inactive Publication Date: 2014-06-04
NAGOYA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the expression of periostin, its presence in the blood, and its relationship with mesothelioma have not been known so far

Method used

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  • Molecular marker for early indentification of pleural mesothelioma patients, and expression analysis method for same
  • Molecular marker for early indentification of pleural mesothelioma patients, and expression analysis method for same
  • Molecular marker for early indentification of pleural mesothelioma patients, and expression analysis method for same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Preparation of anti-periostin antibody

[0073] (1) Structure of human periostin protein

[0074] figure 1 It is a schematic diagram showing the position of the functional domain of the human periostin protein, the structure of each isoform, and the relationship between the structure of the immunogen used for producing the periostin antibody of the present invention. exist figure 1 In, EMI and fascin 1 denote EMILIN homology domain and fascin homology domain, respectively. iso1-4 represent isoforms 1-4 of human periostin, respectively. POSTN781 represents a recombinant protein in which a myc tag and a his tag (denoted "mychis") are fused to the carboxyl terminus of a 781 amino acid polypeptide of human periostin protein isoform 3. POSTN630 represents a recombinant protein in which a myc tag and a his tag are fused to the carboxyl terminus of a polypeptide of 630 amino acids common to all isoforms of human periostin protein comprising up to 4 fascin domains. The ami...

Embodiment 2

[0093] Determination of clinical samples

[0094] (1) Reactivity to clinical samples

[0095] The samples of healthy people were approved by the Ethics Committee of the Institute of Medical Biology of Co., Ltd.; MBL Ethics Review Committee (case number: 022, date of approval: March 27, 2007). Written measurement consent was obtained in advance from each healthy person who provided the sample. The samples of mesothelioma patients were obtained by entrusting BMR (Bio Medical Resources (unified as Sera Care Life Sciences Milford, MA.) at the time of application) (part code: DS-763 description: Mesothelioma Lot#BM203975-BM203986). The above-mentioned plasma sample diluted to 1000 times was used instead of human periostin in the sandwich ELISA method described in Example 1(7) to purify the recombinant protein. Table 2 shows the measurement results of the plasma samples of 16 healthy persons and the plasma samples of 11 mesothelioma patients for the antibody combinations shown in ...

Embodiment 3

[0102] Determination of clinical samples

[0103] (1) Determination of periostin concentration in plasma samples of mesothelioma patients and healthy subjects

[0104] will be used containing 0.1M NaHCO 3 , 0.1M Na 2 CO 3Anti-human periostin monoclonal antibody #6-14-3 diluted to 10 μg / mL with a solution of 0.15% Proclin 150 (SUPELCO), and 50 μL each was coated on a MaxiSorp 96-well plate (NUNC, Thermophysics Corporation, Ltd.), Immobilize the antibody by standing at 4°C overnight or at room temperature for 2 hours. After the antibody solution was removed, 150 μL each of blocking buffer (PBS supplemented with 1% BSA (Proliant, Beritas, Inc.), 0.15% Proclin 150, and 5% sucrose) was dispensed and allowed to stand overnight at 4°C or at room temperature for 2 minutes. Hour. After removing the blocking buffer solution, PBS added with 1% BSA, 0.1% Tween20, 0.15% Proclin150 and 50 μg / mL MAK-33 was used as a diluent to prepare a periostin standard substance (POSTN781 starting fr...

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Abstract

To develop a simple, low-cost and highly reliable testing method for mesothelioma, and a kit used for said testing method. The present invention provides a testing method for mesothelioma comprising a step in which the concentration of human periostin protein is measured in at least one type of sample from among samples of the blood or pleural fluid of a subject. The step in which the concentration of human periostin protein is measured may use an antibody directed against human periostin protein. The present invention further provides a kit for diagnosing mesothelioma, said kit including the antibody directed against human periostin protein. In one kit for diagnosing mesothelioma, the diagnosis of mesothelioma determines if a subject that may have mesothelioma has mesothelioma or is healthy. In another kit for diagnosing mesothelioma, the diagnosis of mesothelioma determines if a subject that may have mesothelioma has mesothelioma or has a respiratory illness other than mesothelioma.

Description

technical field [0001] The present invention relates to a method for examining mesothelioma and a kit for diagnosing mesothelioma. Specifically, it relates to an examination method for mesothelioma comprising a step of measuring the concentration of human periostin (Periostin) in at least one sample of blood and pleural fluid, and a method for mesothelioma comprising an antibody against human periostin protein Kits for tumor diagnosis. Background technique [0002] Mesothelioma is known to occur due to the attraction of asbestos. Mesothelioma is classified according to the site of occurrence into pleural mesothelioma, peritoneal mesothelioma, and peritoneal mesothelioma. Most occur in the pleura, accounting for 80% to 90% of all mesotheliomas (Non-Patent Document 1). Benign pleural mesothelioma is called fibrous mesothelioma. After pneumonia, trauma, etc., fibrous proliferation occurs in the pleura, resulting in a localized mass; in contrast, malignant pleural mesotheliom...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574C07K14/47C07K16/18C12N15/09
CPCC07K14/47C07K16/18G01N33/57407G01N33/57423
Inventor 柳泽圣高桥隆横井香平长谷川好规小野健一郎八木香澄增田瞳竹内俊幸
Owner NAGOYA UNIVERSITY
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