Protein fingerprint method for biological sample analysis

A technology of biological samples and analysis methods, applied in the fields of analyzing materials, biological testing, material separation, etc., can solve the problems of undetectable biomolecules, difficulty in distinguishing biomolecules, complexity, etc.

Inactive Publication Date: 2005-01-19
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 of each dimension (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 between liver cirrhosis and liver cancer caused by hepatitis B

[0064] (1) Experimental method

[0065] Add 1uL of serum to 9M urea, and let the protein react with urea for 30 minutes at 4°C-6°C. Then the above samples were added into the binding solvent (50 mM sodium acetate, pH 4.0-6.0) referred to as BB. Next, the diluted serum was added to the anion adsorbent (anion adsorbent on a silica-coated steel substrate: SO 3 - , that is, anion-exchange adsorbent) on the chip, at room temperature to bind to the chip with anions. Wash twice with BB, then twice with HPLC grade dH2O. Let the chips dry naturally. 0.5 uL of SINAPINIC acid (5 mg / mL 50% acetonitrile; 0.5% trifluoroacetic acid) was added and allowed to dry naturally. directly analyzed by mass spectrometer. Results Mass spectral data were analyzed by computer.

[0066] (2) Experimental results

[0067] Using statistical methods, the form of the analyzed data is a barcode format read...

Embodiment 2

[0077] Example 2 Distinguish between normal and liver cancer

[0078] (1) Experimental method

[0079] Add 1uL of serum to 9M urea, and let the protein react with urea for 30 minutes at 4°C-6°C. Then add the above sample into the binding solvent (50mM sodium acetate, pH4.0~6.0) referred to as BB. Next, add the dilute serum to the anion adsorbent (the anion adsorbent on the steel substrate coated with silicon oxide: SO 3 - , that is, anion-exchange adsorbent) on the chip, at room temperature to bind to the chip with anions. Wash twice with BB, then twice with HPLC grade dH2O. Let the chips dry naturally. 0.5 uL of SINAPINIC acid (5 mg / mL 50% acetonitrile; 0.5% trifluoroacetic acid) was added and allowed to dry naturally. directly analyzed by mass spectrometer. Results Mass spectral data were analyzed by computer.

[0080] Using statistical methods, the form of the analyzed data is a barcode format read by a computer, and the protein fingerprint is displayed as a dark int...

Embodiment 3

[0086] Example 3 Distinguish between normal and liver cirrhosis caused by hepatitis B

[0087] (1) Experimental method

[0088] Add 1uL of serum to 9M urea, and let the protein react with urea for 30 minutes at 4°C-6°C. Then the above samples were added into the binding solvent (50 mM sodium acetate, pH 4.0-6.0) referred to as BB. Next, the diluted serum was added to an anionic adsorbent (anionic adsorbent on a silica-coated steel substrate: SO 3 - , that is, anion-exchange adsorbent) on the chip, at room temperature to bind to the chip with anions. Wash twice with BB, then twice with HPLC grade dH2O. Let the chips dry naturally. 0.5 uL of SINAPINIC acid (5 mg / mL 50% acetonitrile; 0.5% TFA) was added and allowed to dry naturally. directly analyzed by mass spectrometer. Results Mass spectral data were analyzed by computer.

[0089] Using statistical methods, the form of the analyzed data is a barcode format read by a computer, and the protein fingerprint is displayed as...

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PUM

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Abstract

This invention relates to a protein dactylography for analysis of biological sample. 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 method can be used to the diagnosis of cell and clinic diseases, such as tumor with accuracy, convenience and quickness.

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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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/72G01N30/96G01N33/50G01N33/543G01N33/558G01N33/576G06K7/10G06K9/20
Inventor 许洋
Owner 许洋
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