Method for detecting pancreatic cancer

一种检测方法、胰脏癌的技术，应用在检测胰脏癌领域，能够解决不能胰脏癌病人显示出显著的差异等问题，达到快速又简单检测的效果

Inactive Publication Date: 2013-10-09
J OIL MILLS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, assays using these traditional lectins often fail to show significant differences between healthy and pancreatic cancer patients

Method used

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  • Method for detecting pancreatic cancer
  • Method for detecting pancreatic cancer
  • Method for detecting pancreatic cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0127] Examples of the present invention are shown below to illustrate the present invention in detail. However, the present invention is not limited to the following examples.

[0128] (Preparation Example 1) Preparation of PTL

[0129] Agaricus aureus lectin (PTL) was isolated and purified from Agaricus aureus according to the purification procedure shown below.

[0130] (extraction)

[0131] The lyophilized powder (2.5 g) obtained by lyophilizing Agaricus aureus (7.5 g) was extracted with 50 ml, 10 mM Tris buffer (pH 7.2) for 2 hours at 4°C. The obtained extract was centrifuged (15000rpm, 20min, 4°C). Then, the supernatant was filtered through gauze to obtain the first extract. The extraction residue was extracted with 50 ml of 10 mM Tris buffer (pH 7.2) overnight at 4°C. After extraction, centrifuge (15000 rpm, 20 min, 4° C.), and filter the supernatant through gauze to obtain a second extract. Then, these extracts are jointly filtered through filter paper to obtai...

reference example 1

[0146] Reference Example 1: Detection of various glycoproteins in serum by lectin.

[0147] Study by ELISA the affinity of the glycoproteins shown in Table 1 to the biotin-labeled lectins shown below by ELISA:

[0148] PTL (a trehalose α1→6 specific lectin that can be used in the invention), and

[0149] The following commercially available lectins are considered to be specific for trehalose:

[0150] LCA (manufactured by Nippon Seikagaku Co., Ltd., J-Oil Co., Ltd.),

[0151] AAL (manufactured by Nippon Biochemical Industry Co., Ltd., J-Oil Co., Ltd.),

[0152]Lotus (manufactured by Nippon Seikagaku Co., Ltd., J-Oil Co., Ltd.) and

[0153] UEA-I (manufactured by Nippon Seikagaku Co., Ltd., J-Oil Co., Ltd.).

[0154] Serum glycoprotein (human serum albumin (manufactured by CALBIOCHEM), immunoglobulin G (manufactured by Sigma), transferrin (manufactured by Sigma), fibrinogen (manufactured by AbD Serotec), immunoglobulin A (manufactured by BETYL) manufactured), α2-macroglobu...

Embodiment 1

[0167] (Example 1) Lesion haptoglobin in the culture supernatant of pancreatic cancer cell lines was detected by ELISA using PTL.

[0168] Reference Example 1 shows that PTL has no affinity for haptoglobin. Therefore, PTL detects haptoglobin extracted from the medium supernatant of pancreatic cancer cell line (PSN-1, purchased from DS PHARMA) and haptoglobin extracted from human serum as a negative control (purchased from BIODESIGN Company ) were assessed by ELISA. Similar tests to PTL were performed on LCA and AAL as positive controls.

[0169] (Extraction of haptoglobin from pancreatic cancer cells)

[0170] 1000 mL of the culture supernatant of the pancreatic cancer cell line was filtered with an ultrafilter (product name: VIVA SPIN20-10K, manufactured by SARTORIUS), and concentrated to 1 mL. The above concentrated solution was added to the gel (NHS-activated Sepharose 4 fast flow (manufactured by GE Healthcare)) in which the anti-haptoglobin antibody (manufactured by Th...

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Abstract

To provide a method of accurately detecting pathological haptoglobin using a lectin having strong affinity and high specificity for fucose. The method of the present invention for detecting pancreatic cancer is characterized in that a fucose ±1†’6 specific lectin is allowed to act on pathological haptoglobin contained in a sample obtained from a living body, said lectin: (1) being extractede from basidiomycetes, (2) having a molecular weight of 4,000 to 40,000 as determined by the SDS polyacrylamide gel electrophoresis, and (3) having affinity for a fucose ±1†’6 sugar chain with a binding constant of 1.0 × 10 4 M -1 or more at 25°C.

Description

technical field [0001] The present invention relates to a method for detecting pancreatic cancer, more specifically, to a method for detecting pancreatic cancer by using lesion haptoglobin as a tumor marker. Background technique [0002] Because the pancreas is located deep in the center of the body, cancerous changes in the pancreas are difficult to detect. Tumor markers used to diagnose pancreatic cancer include CEA (standard value: 5.0 ng / mL) and CA 19-9 (standard value: 37 U / mL). However, false positives are also included in the results from these tumor markers, so tumor markers alone may not give reliable results. In order to confirm the diagnosis of pancreatic cancer, costly and thorough examinations such as: computerized tomography (CT), endoscopic retrograde cholangiopancreatography (ERCP), endoscopic ultrasonography (EUS) and angiography are necessary. Among them, EPCR and EUS are invasive, which may increase the burden on patients. [0003] Recent reports ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N2333/375G01N33/57438G01N2400/00G01N33/574G01N33/68
Inventor 小林夕香草间健龟井麻直
Owner J OIL MILLS INC
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