Use of detecting urine protein fingerprint by protein fingerprint technique

A technology of fingerprinting and urine, which is 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, and difficulty in distinguishing biomolecules

Inactive Publication Date: 2005-08-24
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
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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  • Use of detecting urine protein fingerprint by protein fingerprint technique
  • Use of detecting urine protein fingerprint by protein fingerprint technique

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1 Normal people, nephrotic syndrome and lupus erythematosus nephritis in the distinction of protein fingerprint in urine

[0063] (1) Experimental method

[0064] 1. Materials

[0065] 1. Source of specimens: There were 36 nephrotic syndrome patients in the nephrotic syndrome group. There were 30 patients with lupus erythematosus erythematosus in the lupus erythematosus nephritis group. There were 32 normal people in the normal group.

[0066] 2. Reagents: urea, acetonitrile, trifluoroacetic acid, and SPA (Sinapinic acid) were all purchased from Sigma.

[0067] 2. Method

[0068] 1. Collection of samples: Urine was collected in the middle of the morning.

[0069] 2. Sample preparation: take 100 μl of urine, centrifuge at 14,000 rpm for 5 minutes, absorb the supernatant, and store at -70°C.

[0070] 3. Chip pretreatment: Take 5 μl of urine supernatant and spot on the hydrophobic chip, incubate in a wet box at room temperature for 30 minutes, wash the spo...

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Abstract

The invention relates to be detained the chromatographic analysis law capture protein group to carry on the analysis protein fingerprint system. This method carries on the distinction examination protein group with the mass spectrography. Analyzes the obtained data the form is may use the bar code form which the computer machine reads (protein fingerprint). This invention method may use in examines in the urine the normal person and the patient (for example nephrosis synthesis card and lupus erythematosus nephritis) the urine protein fingerprint. This method accurate, is convenient also quickly.

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): G01N30/90
Inventor许洋
Owner许洋