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Use of protein fingerprint method in detecting protein fingerprint of pancreas cancer cell in blood

A protein and protein technology, applied in the field of protein fingerprinting and protein fingerprinting of biological sample analysis, can solve the problems of inability to observe molecular weight, inability to detect biomolecules, complexity, etc.

Inactive Publication Date: 2005-03-16
BEIJING SELDI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First, the method provides only two characteristics of biomolecules - mass and isoelectric point (pI)
Second, the resolution in each dimension is limited by the resolving power of the gel
For example, it is often difficult to distinguish between biomolecules that differ by less than 5% in mass or that differ in pI
Third, gels have limited capacity and sensitivity and may not be able to detect small amounts of expressed biomolecules
Fourth, small proteins or peptides with a molecular weight below about 10-20 kDa cannot be observed
Since human cell proteins are much more complex than potatoes, electrophoresis alone to identify protein fingerprints of diseases is far from meeting clinical needs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Differentiation of protein fingerprints in blood between normal human cells and pancreatic cancer patients

[0064] (1) Experimental method

[0065] 1. Material

[0066] 1. Specimen source: There are 33 cases in the pancreatic cancer patient group, all of which were pathologically diagnosed as pancreatic cancer. There are 33 cases in the normal group (healthy volunteers with normal health checkup).

[0067] 2. Reagents: Urea, acetonitrile, trifluoroacetic acid, and SPA (Sinapinic acid) were all purchased from Sigma.

[0068] Two, method

[0069] 1. Sample collection: After collecting whole blood, draw serum and store at -80℃;

[0070] 2. Sample preparation: Take 20μl of serum and dilute it to 150μl with pH7.0 buffer (100mmol / L sodium phosphate, 250mmol / L sodium chloride) according to 1:7.

[0071] 3. Chip pretreatment: Coordinated covalent metal chelating agent acts as adsorbent chip. Add 50μl of 100mmol / L copper sulfate to each hole, shake for 5min at room temper...

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PUM

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Abstract

This invention also relates to protein dactylography method for analyzing for the protein group got by retention chromatography. This method adopts mass spectroscopy to distinguish and detect protein group. The analysis data takes the form of bar code (protein dactylography) read by computer. This invention method can be used to detect protein fingerprint in the normal cell and cancer of stomach patient cell in the blood.

Description

Technical field [0001] The present invention relates to a new protein analysis method in biological samples, and specifically refers to the protein fingerprint method for the analysis of biological samples. The present invention also relates to protein fingerprinting methods in which proteomes are captured using retention chromatography and analyzed in a computer-readable barcode format (protein fingerprinting). Background technique [0002] Both the normal functions and pathological characteristics of cells depend to a certain extent on the functions of proteins expressed by cells. Therefore, identifying differences in proteins expressed within cells can be used to diagnose disease and ultimately for drug development and disease treatment. To carry out differential analysis of protein expression and function, it is required to be able to resolve complex mixtures of intracellular molecules. However, many substances in cells often exist in trace amounts. The current methods...

Claims

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Application Information

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IPC IPC(8): G01N30/00G01N30/72G01N33/53G01N33/68G06K7/10H01J49/04
Inventor 许洋陈国强
Owner BEIJING SELDI TECH
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