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Method for analyzing proteome in biological sample

A biological sample and analysis method technology, applied in the preparation of test samples, analytical materials, biological testing, etc., can solve the problems of limited resolution, difficult to distinguish, limited gel capacity and sensitivity, etc., to improve the detection rate , early diagnosis of cancer, the effect of increasing the difference

Active Publication Date: 2010-03-31
苏州云泰生物医药科技有限公司 +1
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AI Technical Summary

Problems solved by technology

Firstly, this method can only distinguish protein molecules based on their molecular mass and isoelectric point (pI); secondly, the resolution of each dimension is limited by the resolving power of the gel, and it is usually difficult to distinguish between those with a mass difference of less than 5% or isoelectric points (pI). similar biomolecules
Third, the capacity and sensitivity of gels are limited, and small and micro-expressed biomolecules may not be detected
Fourth, small peptides with a molecular weight below about 10-20 kDa cannot be observed
[0011] However, due to the design of the reaction conditions of the above patents, the sensitivity, that is, the early detection rate, still needs to be improved.

Method used

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  • Method for analyzing proteome in biological sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Diagnosis of liver cancer

[0041] 1. Establish a standard fingerprint library of normal serum, the method is as follows:

[0042] 1,000 healthy volunteers with three years of continuous physical examination, half male and half male, aged 20 to 50, no family history, at least two generations of cancer-free patients, a few drops of blood were drawn, and the serum was separated. Add 1 μL of serum to 2 times the volume of U9 buffer to dilute (containing 9mol / L urea, 2% CHAPS, 50mmol / L Tris hydrochloric acid, 1% DTT, pH9.0), shake in ice bath for 30min, and make the proteome interact with urea. Then, 3 μL of 40 mM sodium acetate binding reagent at pH 3.75 was added to the above sample; MACS nano-magnetic beads (product of Miltenyi Biotechnology Co., Ltd., Germany) were added to the serum, and incubated on a magnet for 10-30 min; the supernatant was removed; Then add the binding reagent, place it on the magnet and incubate for 1-5 minutes to make the nano-magnetic...

Embodiment 2

[0057] The diagnosis of embodiment 2 gastric cancer

[0058] 1. Establish a standard fingerprint library of normal serum:

[0059] 1,000 healthy volunteers with three consecutive years of physical examination, half male and half male, aged 20 to 50, no family medical history, at least two generations of cancer-free patients, a few drops of blood were drawn, and treated according to the method of Example 2. The difference is: the pH of the binding reagent is 3.95.

[0060] 2. Establish a serum fingerprint database for gastric cancer;

[0061] 52 patients were diagnosed with gastric cancer by CT, MRI and other examinations, half male and half male, aged 40 to 60, all patients underwent surgical resection of the tumor, and the pathological results proved to be gastric cancer. After the whole blood was collected before the operation, it was processed according to the method under the sample collection item, and the serum was collected and stored at -80°C; it was processed accord...

Embodiment 3

[0065] Example 3 Diagnosis of rectal cancer by body fluid

[0066] 1. Establish a standard fingerprint library of normal urine

[0067] A total of 1,000 healthy volunteers with three consecutive years of physical examination, half male and half male, aged 20 to 50, no family medical history, no cancer patients for at least two generations. The collected urine was processed and analyzed by mass spectrometry according to the steps in Example 2, except that the pH of the binding reagent used was 3.55.

[0068] 2. Establish a urine fingerprint database for rectal cancer;

[0069] CT, MRI and other examinations confirmed rectal cancer in 278 patients, half male and half male, aged 40 to 60. All patients underwent surgical resection of the tumor, and the pathological results proved to be rectal cancer. The collected urine was processed and analyzed by mass spectrometry according to the steps in Example 1, except that the pH of the binding reagent used was 3.55.

[0070] 3. Compar...

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Abstract

The invention aims to provide a method for analyzing proteome in a biological sample. The method can detect unique proteome generated by cancer cells through blood or body fluid in an early stage of tumors; and the method can diagnose cancers more sensitively and more early so as to make early diagnosis and treatment of the cancers possible.

Description

technical field [0001] The present invention relates to a method for proteome analysis in biological samples, in particular to an analysis method for identifying specifically expressed proteomes in different biological samples, more precisely, a method for identifying specifically expressed proteins produced by tumor cells, belonging to the biological field of medicine. Background technique [0002] At present, in large and medium-sized cities, among the cancer patients seen in the outpatient department of the hospital, generally speaking, the early (stage I) patients account for about 5% to 10%, the middle stage (II stage) accounts for about 20%, and the patients in the later stage (III + IV stage) accounted for about 70% to 75%. The 5-year survival rate of advanced patients after treatment is only 10% to 30%. From the 1960s to the present, some medical personnel and scientific researchers in my country have carried out preliminary screening, general survey and general tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62G01N1/28G01N33/68G01N33/574
Inventor 高尚先
Owner 苏州云泰生物医药科技有限公司
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