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Microflow control chip for protein analysis and its application in protein analysis

A microfluidic chip and protein technology, which is applied in the direction of analytical materials, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of unfavorable proteome, non-universal, cumbersome process, etc., and achieve fast and convenient analysis Effect

Inactive Publication Date: 2004-01-21
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Connecting the two for protein two-dimensional capillary electrophoresis is the best analysis method, but connecting two capillary columns requires professional operation, which is not universal, and is very unfavorable for proteome research
[0006] The currently used protein analysis methods are mainly to separate and analyze protein samples to obtain one-dimensional information of the samples. To obtain more information, a combination of various analysis methods is required. The collected protein is hydrolyzed by protease, mainly using specific protease, and the obtained hydrolyzate (mainly peptide fragments) is separated and analyzed by high performance liquid chromatography / mass spectrometry, capillary electrophoresis / mass spectrometry, and mass spectrometry, so that The process is cumbersome, and many processes cannot be automated, and it takes a long time

Method used

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  • Microflow control chip for protein analysis and its application in protein analysis
  • Microflow control chip for protein analysis and its application in protein analysis
  • Microflow control chip for protein analysis and its application in protein analysis

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Taking hen ovalbumin as an example, the protein has a molecular weight of 43kDa and consists of 386 amino acids. The chips used are figure 1 The designed chip is made of quartz, and the channel size is 50 μm × 20 μm. The channels in the first and second units are modified with linear polyacrylamide, and in the channel in the third unit, the channel in the direction from the cross to the outlet is decorated with Trypsin was modified. 100mg / mL hen egg protein is being analyzed in the first unit, the injection channel is 0.5 cm, the separation channel is 2 cm, and the buffer solution used is 20 mmol / L sodium hydroxide and 4% methylcellulose solution as the cathode Electrolyte, 100mmol / L phosphoric acid is used as the anode electrolyte, the sample injection electric field is 200V / cm, the separation electric field is 260V / cm, the time takes about 60 seconds; then part of the samples enter the second unit under the action of the electric field, and the second unit The injec...

Embodiment 2

[0027] Using turkey ovalbumin as a sample, the protein has a molecular weight of 43kDa and is also composed of 386 amino acids, 36 of which are different from hen ovalbumin. The chips used are figure 1The designed chip is made of quartz, and the channel size is 50 μm × 20 μm. The channels in the first and second units are modified with linear polyacrylamide, and in the channel in the third unit, the channel in the direction from the cross to the outlet is decorated with Trypsin was modified. 100mg / mL turkey egg protein is being analyzed in the first unit, the injection channel is 0.5 cm, the separation channel is 2 cm, and the buffer solution used is 20 mmol / L sodium hydroxide and 4% methylcellulose solution as the cathode Electrolyte, 100mmol / L phosphoric acid is used as the anode electrolyte, the sample injection electric field is 200V / cm, the separation electric field is 260V / cm, the time takes about 60 seconds; then part of the samples enter the second unit under the acti...

Embodiment 3

[0029] Taking the mixture of bovine serum albumin, transferrin and concanavalin A as samples, their isoelectric points are 5.82, 7.31 and 5.42 respectively; molecular weights are 69.3kDa, 35.8kDa and 10.6kDa respectively. The sample is 200mg / L, and the chip used is according to figure 1 The designed chip is made of quartz, and the channel size is 50 μm × 20 μm. The channels in the first and second units are decorated with linear polyacrylamide, and the channels in the third unit are on the channel in the direction from the cross to the outlet. Modification with trypsin. In the first unit, the sampling channel is 0.5 cm, the separation channel is 4 cm, 20 mmol / L sodium hydroxide and 4% methylcellulose solution are used as the catholyte, and 100 mmol / L phosphoric acid is used as the anode electrolyte to form an electrophoretic gradient. is 600V / cm, the injection electric field is 300V / cm, and the separation electric field is 400V / cm, transferrin and the other two proteins can b...

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Abstract

The microflow control chip for protein analysis features that it consists of four basic units connected successively with the sample outlet of the upper basic unit being connected to the sample inletof the next basic unit. They are the first protein isoelectric focusing separation unit; the second protein screening electrophoresis unit with organic polymer selected from cellulose and agarose asscreening medium; the third protein proteinase enzymolyzing unit with specific proteinase bonded to the inner wall or specific proteinase stuffing filled; and the fourth interface unit to convey the enzymolysis product of protein to mass spectral analysis. Using the chip of the present invention in protein analysis can obtain great amount of information in short time and through simple operation.

Description

Technical field: [0001] The invention relates to protein analysis technology, and in particular provides a microfluidic chip specially used for protein analysis. Background technique: [0002] The study of proteome is one of the basic tasks in the post-genome era. The development of proteomics research relies more on the development of analytical methods. Currently, the main technologies used for protein analysis are two-dimensional gel electrophoresis, computer image analysis, protein identification technology, high performance liquid chromatography, and capillary electrophoresis. The most widely used method in biology is two-dimensional gel electrophoresis. Although this method is relatively mature, the entire analysis process is cumbersome and time-consuming due to the inability to perform automated operations and analysis under high electric fields due to technical reasons. , and this method is not quantitatively accurate. [0003] Protein identification techniques mai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/37C12Q1/68G01N33/50G01N33/53G01N33/543G01N33/68
Inventor 毛秀丽王克夷林炳承
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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