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Use of enhanced protein fingerprint spectrum

A protein and fingerprint technology, applied in the field of protein fingerprinting and protein fingerprinting of biological sample analysis, can solve problems such as difficulty in distinguishing biomolecules, limited gel capacity and sensitivity, and inability to detect biomolecules.

Inactive Publication Date: 2006-10-11
许洋
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  • Summary
  • Abstract
  • Description
  • Claims
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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

[0071] Example 1 Sample Tests of Normal Human Samples and Cancer Patient Groups

[0072] (1) Experimental method

[0073] 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 2 O 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.

[007...

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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. Said bar code can be transformed into one strengthen protein fingerprint drawing instructed by standard mass control serum; the protein can be kept by bar code (digital code) or drawing (strength protein fingerprint drawing) forever after being detected. The invention can be used to detect the protein fingerprint of normal cell and sufferer 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). Its barcode format can be converted into an enhanced protein fingerprint (Enhanced Proteomic Graphics; EnhanProGram or EPG) under the guidance of standardized quality control serum. 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 abilit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/90G01N30/95G01N30/92
Inventor 许洋
Owner 许洋
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