Application of high expression or low expression of modified protein to tumor cell detection

A protein and protein technology, applied in the field of protein fingerprinting and protein fingerprinting in the analysis of biological samples, can solve the problems that it is difficult to distinguish biomolecules, protein cancer cell markers are not reported, and biomolecules cannot be detected.

Inactive Publication Date: 2006-10-11
许洋
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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
However, there is no report on protein modifications (methylation, acetylation, hydroxylation, phosphorylation, etc.) as cancer cell markers

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1 Sample Tests of Normal Human Samples and Cancer Patient Groups

[0069] (1) Experimental method

[0070] Add 1 μL of serum to 9M urea, and let the protein react with urea for 30 minutes at 4oC-6oC. Then the above samples were added into the binding solvent (50 mM sodium acetate, pH 4.0-6.0). Next, the diluted serum was added to an anionic adsorbent (anionic adsorbent on a silica-coated steel substrate: SO 3 - , that is, on the matrix of anion-exchange adsorbent), it is combined with an anion-bearing matrix at room temperature. Wash twice with binding solvent followed by HPLC grade dH 2O wash twice. Let the substrate dry naturally. Add 0.5 μL of Sinapinic acid (5 mg / mL 50% acetonitrile; 0.5% trifluoroacetic acid) and let it dry naturally. First use the mass spectrometer to standardize the quality control serum to adjust the zero point, and then directly analyze it with the mass spectrometer. Results Mass spectral data were analyzed by computer.

[0071...

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Abstract

The invention discloses a protein fingerprint method for analyzing the protein group. It uses mass spectrography to detect the protein group, and the usable data analyzed by statistics is the bar cod (protein fingerprint) red by the computer. The invention can be used to detect the decorate protein fingerprint of tumor cell, with high speed and simple process.

Description

technical field [0001] The invention relates to a novel method for analyzing proteins in biological samples, in particular to a protein fingerprint method for analyzing biological samples. The present invention also relates to protein fingerprinting in which the proteome is captured by retention chromatography and analyzed using a computer readable barcode format (protein fingerprint). Background technique [0002] Both normal function and pathological characteristics of cells depend to some extent on the function of proteins expressed by cells. Therefore, identifying differences in proteins expressed within cells can be used in the diagnosis of disease and ultimately in drug development and disease treatment. However, differential analysis of protein expression and function requires the ability to resolve complex mixtures of intracellular molecules. However, many substances in cells often exist in trace amounts. The current methods for analyzing proteins have limitations ...

Claims

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Application Information

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IPC IPC(8): G01N30/90G01N30/95G01N30/92
Inventor许洋
Owner许洋