Method for detecting four kinds of tumor serum proteins

A protein and tumor technology, applied in the field of four common tumor serum proteins of liver cancer, breast cancer, meningioma, and brain glioma detection, which can solve the difficulties of early molecular detection of tumors, evaluation of efficacy and prognosis, recurrence and metastasis monitoring and biological treatment. And other issues

Inactive Publication Date: 2005-03-02
ZHEJIANG UNIV
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  • Abstract
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Problems solved by technology

This situation has caused certain difficulties in early molecular detection of tumors, evaluation of curative effect and prognosis, monitoring of recurrence and metastasis, and biological treatment.

Method used

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  • Method for detecting four kinds of tumor serum proteins
  • Method for detecting four kinds of tumor serum proteins
  • Method for detecting four kinds of tumor serum proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 Detection method of the present invention

[0023] 1. Serum samples were thawed in an ice bath and centrifuged at 3000rpm for 5 minutes, and 10ul was diluted with 90ul 0.5% CHAPS (3-cycloethylamine-1-propanesulfonic acid) (pH7.4) solution and then added to Cibacron Blue3GA (white In the protein affinity resin Sigma), react on a shaker at 4°C for 60 minutes, centrifuge at 12000rpm for 5 minutes (4°C), take 50ul of the supernatant solution, and further use 20mM HEPES [N-(2-hydroxyethyl)-piperazine- N'-2 (propanesulfonic acid)] (pH7.4) (H4) / 50mM sodium acetate (pH4.0) (WCX2) was diluted to 200ul, added to the Bioprocessor (96-well sample loaded with H4 / WCX2 chip) (Ciphergen, USA) and vibrated on a shaker for 60 minutes. Shake off the serum specimen, wash the chip with 200ul of 20mM HEPES (pH7.4) (H4) / 50mM sodium acetate (pH4.0) (WCX2) 3 times x 5 minutes, then wash the chip 2 times x 1 minute with deionized water, and dry Afterwards, 0.5ul energy absorbing m...

Embodiment 2

[0036] Example 2 Brain Tumor Detection

Embodiment 3

[0087] Example 3 Breast cancer detection

[0088] Using the method in Example 1, the serum protein fingerprints of 49 breast cancer patients, 37 breast cancer patients and 33 healthy people were detected and analyzed. List all the differentially expressed protein peaks obtained in the comparison between breast cancer and healthy human serum protein mass spectrometry data, and use the method of gradually increasing the number of mass-to-charge ratio peaks according to the relative importance to establish multiple comprehensive neural networks and evaluate their prediction success rate, and then The first four mass-to-charge ratio peak combinations with the highest detection accuracy were screened out, and their mass-to-charge ratios were 17.3kDa, 26.3kDa, 5.7kDa and 8.9kDa, respectively. In all 82 specimens, 55 cases (32 cases of breast cancer patients and 23 cases of healthy people) were re-randomly sampled to establish a new training set, and the mass spectrometry data of pro...

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Abstract

The invention provides a detecting method for the blood serum protein of four kind of familiar tumours which are, tumour, meningioma tumour, mammary tumour and hepatic tumour by the following steps: (1) using the surface concentrated laser SELDI to determine the protein group graphoes of the blood serum specimen of the tumour patient and the healthy person. (2) screening the corresponding tumour marker and establishing the detecting model for analysis integrated the biological informatics technique. The method of the invention is adopted in the process of selecting the tumour medicine and provides a new way and method for early discovery and detection of the tumour. The discovered new seriial protein m/z peak also provides a base and resource for seeking new better tumour marker, meanwhile, provides a new thought for exploring the generation and development of tumours.

Description

technical field [0001] The invention relates to the field of early detection of tumors, in particular to a method for in vitro detection of four common tumor serum proteins of glioma, meningioma, breast cancer and liver cancer, which can be used in screening these common tumor therapeutic drugs. It also provides a series of candidate tumor markers for further research. Background technique [0002] With the change of human disease spectrum, tumor has become one of the common causes of death and an important public health problem that seriously endangers human health. At present, how to improve the detection rate of tumors, especially the early detection rate, has been recognized as the key to improving its curative effect and prognosis. With the smooth implementation of the Human Genome Project, functional genomics with protein as the main research object has gradually become one of the most dynamic fields in life science. Since protein, not nucleic acid, is the specific e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/72G01N30/86G01N30/88G01N33/48
Inventor 郑树余捷凯刘建胡跃张书展
Owner ZHEJIANG UNIV
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