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Substrate for labo-on-a-chip

A lab-on-a-chip and substrate technology, which can be used in material inspection products, measuring devices, and material analysis through electromagnetic means, and can solve problems such as weak binding between polyalkylene glycol and substrates, and inability to separate and swim proteins.

Inactive Publication Date: 2007-04-04
TORAY IND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in this method, the bonding force between polyalkylene glycol and the substrate is weak, and when washing with a solvent, a relatively large amount of polyalkylene glycol comes off.
Therefore, only electrostatically coating polyalkylene glycol on the channel wall surface of a chip having micro-channels made of a polymer substrate cannot separate and migrate proteins.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] A polymethyl methacrylate substrate having a size of 20×60 mm and a thickness of 0.2 mm was immersed in an aqueous polyethylene glycol solution having a molecular weight of 500,000 and 2,000 ppm. The dipped polymethyl methacrylate plate was sealed and irradiated with 2.5 kGy of gamma rays to perform grafting. The substrate irradiated with γ-rays was dried, and attached to a fluorescent plate having holes in the light-transmitting portion. On the attached fluorescent plate, an aqueous solution of FITC-labeled BSA protein and IgG protein diluted to 10 μg / ml was solidified at room temperature for 10 minutes, and after removing the solution, it was washed with phosphate buffer (PBS) to measure fluorescence. strength.

Embodiment 2

[0100] Microvias of a polymethacrylate substrate having microvias of width 100 μm x depth 60 μm x length 50 cm were filled with aqueous polyethylene glycol solutions with molecular weights of 500,000 and 2,000 ppm, and 2.5 kGy of γ-rays were irradiated to perform grafting. After irradiation, the polyethylene glycol aqueous solution in the microchannel was taken out and washed with purified water. After injecting the cell-free protein synthesis reaction solution from Escherichia coli into the microchannel, place it at 30°C for 1 hour to synthesize chloramphenicol acetyltransferase (CAT), which converts acetyl from Acetyl CoA transfers to the 3'-hydroxyl of chloramphenicol, 26,000 molecular weight enzyme. The CAT protein synthesized in the microchannel was recovered and quantified by ELISA as Example 2.

Embodiment 3

[0108] A chip for electrophoresis made of polymethyl methacrylate and having channels with a diameter of 100 μm was immersed in an aqueous polyethylene glycol solution having a molecular weight of 500,000 and 2,000 ppm. The dipped chip for electrophoresis was sealed and irradiated with 2.5 kGy of γ-rays to perform grafting. Remove the polyethylene glycol in the channel, fill with 5% polyacrylamide (molecular weight 600,000-1,000,000) 0.1M Tris-aspartic acid (pH8) solution, add 5% to parts A, B, and C of Figure 3 Polyacrylamide (molecular weight: 600,000-1,000,000) 0.1M Tris-aspartic acid (pH8) solution, add fluorescently labeled trypsin inhibitor and fluorescently labeled BSA to 0.05MTris-HCl containing 1% SDS in part D ( pH8) solution. Electrodes were inserted into parts A, B, C and D of the electrophoresis chip filled with polyacrylamide and protein solution, and a voltage of 350 V was applied to part B for 1 minute. Afterwards, 500V was applied to C, and 150V was applied ...

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Abstract

A substrate for labo-on-a-chip comprising a resin base material and hydrophilic polymer bonded covalently to the surface thereof with a high energy beam, especially a protein treatment chip. A substrate for labo-on-a-chip capable of long time use while exhibiting cleaning resistant effect in which protein is not adsorbed to the surface of the base material can be provided. Since detection noise is reduced, a trace of protein can be analyzed with high accuracy and a polymer chip for protein electrophoresis having a microchannel can be provided.

Description

technical field [0001] The present invention relates to a substrate for a lab-on-a-chip for performing flow or reaction or analysis of a protein solution in a device or facility for structural and functional analysis of proteins and reactions using proteins . Background technique [0002] At present, a chip including microchannels for carrying out various chemical reactions is a technology that is gradually attracting attention from the viewpoint of reaction efficiency, speed, and various reagents. In general, the concept of a new analytical method for performing chemical reactions and analyzes in microchannels formed on a glass chip of several centimeters square, which has been called a "lab-on-a-chip", has been gradually recognized. In the future, with the development of biotechnology, the use of microchannels will become an indispensable technology in the field of biochemistry, and the application of microchannels to protein structure and function analysis and reactions ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447G01N33/68
CPCG01N27/44747G01N27/44756
Inventor 山崎义昭川添直辉日笠雅史郑基晚信正均村上裕二
Owner TORAY IND INC
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