Application of polypeptide tagging object in detecting tumor in large intestine, and monitoring after operation

A technology for large intestine and tumor, applied in the application field of polypeptide markers, can solve the problems of unclear protein function and large research space, and achieve the effect of improving the level of diagnosis and treatment and postoperative survival rate

Inactive Publication Date: 2007-07-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] What kind of proteins are the vast majority of new candidate markers discovered by SELDI mass spectrometry and their role in the development of tumors is still unclear, and there is a lot of room for research

Method used

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  • Application of polypeptide tagging object in detecting tumor in large intestine, and monitoring after operation
  • Application of polypeptide tagging object in detecting tumor in large intestine, and monitoring after operation
  • Application of polypeptide tagging object in detecting tumor in large intestine, and monitoring after operation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Identification of colorectal cancer-related proteins

[0023] The sample used for separation is a 1ml serum sample from a patient with colorectal cancer. Serum was initially separated by SPE column (reversed phase, acetonitrile gradient) (monitored by SELDI-TOF-MS for the eluted fraction where the target protein was located), and then enriched by a freeze-dryer. After redissolving, use high-performance liquid chromatography-mass spectrometry (HPLC-MS) to separate and purify (use MALDI-TOF-MS to monitor the elution component where the target protein is located), and further digest the target elution component with trypsin. Several of the fragments were sequenced by LC / MS / MS, the amino acid sequence was determined and submitted to MASCOT (MatrixScience, www.matrixscience.com) to determine the source of the target protein peak. After determining the primary sequence of the protein, purchase the corresponding antibody, and use the antibody to bind to the high-ex...

Embodiment 2

[0031] Example 2 Differential expression of colorectal cancer-associated polypeptide 1 in colorectal cancer and control samples

[0032] Serum samples from 55 cases of colorectal cancer and 92 healthy people were combined with H4 chip and detected by SELDI mass spectrometer. Further use the CM10 chip to detect 124 cases of colorectal cancer samples and 46 healthy controls to verify the difference of colorectal cancer-related polypeptide 1

[0033] 1 Chip pretreatment

[0034] l Put the chip into the 96-well protein chip working bracket Bio-processor, and write down the chip number.

[0035] lAdd 200μl Binding Buffer corresponding to the chip to each well:

[0036] H4: 20mM HEPES (pH=7.4)

[0037] CM10: 50mM sodium acetate (pH=4.0)

[0038] l Oscillate on a shaker (600rpm, room temperature) for 5 minutes to fully balance, remove the liquid, and repeat the balance once.

[0039] 2 Pretreatment of serum

[0040] l Melt the serum slowly on an ice bath (30-60 minutes), and ce...

Embodiment 3

[0058] Example 3 Application of colorectal cancer-associated polypeptide 1 in postoperative detection and prognosis estimation of colorectal cancer

[0059] 1. Materials and Methods

[0060] 1.1 Materials

[0061] Serum samples from 10 patients with colorectal cancer before operation, 3 months after operation, 6 months after operation, 12 months after operation, and 18 months after operation.

[0062] Protein Chip Selection: H4 Hydrophobic Surface Chip.

[0063] 1.2 Experimental method

[0064] 1.2.1 According to the standard of diagnosis and treatment of colorectal cancer, the serum CEA, hepatobiliary color Doppler ultrasound or CT, bone ECT, chest X-ray, and electronic colonoscopy were reexamined before and after 3 months, 6 months, 12 months, 18 months, and 24 months. Establish a database of clinical and pathological data, postoperative treatment and follow-up data of colorectal cancer based on ACCESS.

[0065] 1.2.2 The experimental procedure of serum protein mass spec...

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Abstract

An application of polypeptide mark object in large intestine cancer detection includes knowing that correlation polypeptide content in serum specimen of large intestine cancer is obviously raised, using surface intensified laser de-absorption ionization flying time mass analyzer to determine correlation polypeptide content in serum specimen of patient for confirming whether detected patient is large intestine cancer patient or not.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of a polypeptide marker, in particular to the application of a polypeptide marker that can be used for early detection, postoperative monitoring and prognosis evaluation of colorectal tumors. Background technique [0002] Colorectal cancer is one of the complex human diseases and one of the nine common malignant tumors in my country. According to the statistical results recently published by the National Cancer Prevention and Control Office, the adjusted mortality rate of colorectal cancer is listed as the fifth major malignant tumor, and the incidence rate in big cities is the third for men and the second for women. Although the diagnosis and treatment of colorectal cancer have made great progress in recent years, the overall curative effect of colorectal cancer has not been significantly improved, and the 5-year survival rate after surgery is still hovering around 50%...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/52G01N23/225
Inventor 余捷凯陈益定郑树徐文鸿张舒展
Owner ZHEJIANG UNIV
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