Molecular markers and their expression analysis methods for early detection of patients with pleural mesothelioma

一种患者、皮瘤的技术，应用在间皮瘤的检查领域，能够解决骨膜蛋白间皮瘤关系未知等问题

Inactive Publication Date: 2016-05-04
NAGOYA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 markers and their expression analysis methods for early detection of patients with pleural mesothelioma
  • Molecular markers and their expression analysis methods for early detection of patients with pleural mesothelioma
  • Molecular markers and their expression analysis methods for early detection of patients with pleural mesothelioma

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 commissioned by BMR (BioMedicalResources (unified as SeraCareLifeSciencesMilford, MA.) at the time of application) (part code: DS-763 description: MesotheliomaLot#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 Table 1.

[0...

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 , 0.1MNa 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 fro...

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Abstract

An object of the present invention is to develop a simple, inexpensive, and highly reliable method for examining mesothelioma, and a kit used in the method. As a solution to the problem, the present invention provides a method for examining mesothelioma, comprising the step of measuring the concentration of human periostin protein in at least one sample of blood or pleural fluid of a subject. In some cases, the step of measuring the concentration of the human periostin protein uses an antibody against the human periostin protein. The present invention provides a kit for the diagnosis of mesothelioma, comprising an antibody against human periostin protein. In the kit for diagnosing mesothelioma of the present invention, the diagnosis of mesothelioma may be to determine whether a subject who may have mesothelioma is a mesothelioma patient or a normal person. In the kit for diagnosing mesothelioma of the present invention, the diagnosis of mesothelioma may be to determine whether a subject who may suffer from mesothelioma is a patient with mesothelioma or a respiratory organ disease other than mesothelioma of patients.

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 Patents(China)
IPC IPC(8): G01N33/574C07K14/47C07K16/18C12N15/09
CPCC07K16/18G01N33/57407C07K14/47G01N33/57423
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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