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Method for fixing protein on surface of protein chip

A protein chip and surface immobilization technology, applied in the field of immobilizing proteins on the surface of protein chips, can solve problems such as protein instability, low protein biological activity, protein spatial conformation changes, etc., to inhibit physical adsorption, reduce interaction, and reduce impact. Effect

Inactive Publication Date: 2006-10-25
INST OF MECHANICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the defects that the existing methods for immobilizing proteins cause large changes in protein spatial conformation, low protein biological activity, and unstable protein fixation, thereby providing a protein that is stable and stable in spatial conformation. A method for immobilizing proteins on the surface of protein chips with little impact and high biological activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Prepare a mixed ethanol solution of methyltriethoxysilane and aminopropyltriethoxysilane (the molar ratio of the two is 20:1), and the concentration of methyltriethoxysilane is 1 mmol;

[0026] Put the protein chip with the silicon chip as the substrate in the above solution and soak for 5 hours for modification; wash the solid substrate with absolute ethanol for 3 times; then use glutaraldehyde to activate the amino group on the aminopropyltriethoxysilane ;

[0027] Prepare a 1 mg / ml human immunoglobulin G PBS (phosphate buffer) solution containing 0.05 wt% Tween 20; put the modified protein chip into the human immunoglobulin G solution and soak for 30 minutes.

Embodiment 2

[0029] Prepare the mixed ethanol solution of undecanethiol and carboxyl undecanethiol (the molar ratio of the two is 100:1), the concentration of undecanethiol is 1 mole;

[0030] Place the protein chip with gold as the substrate in the above solution and soak for 30 minutes for modification; wash the solid substrate with absolute ethanol for 3 times; then use ethyl carbodiimide (EDC) to activate carboxyundecanethiol carboxyl group;

[0031] Prepare 1mg / ml human serum albumin PBS solution, which contains 1wt% Tween 20;

[0032] Put the modified protein chip into the human serum albumin solution and soak for 30 minutes.

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Abstract

The invention relates to a method for immobilizing proteins on the surface of a protein chip. The method uses the dehydrated ethanol solution of component A and component B to soak the protein chip, and the component A is an organic compound that can make the surface of the protein chip hydrophobic; The protein molecule is covalently linked with an active group, an organic compound containing a polar group at one end; then use conventional methods to activate the polar group of component B; finally put this protein chip into the protein solution to be immobilized soak. The protein immobilized by this method is relatively stable; and it can effectively reduce the interaction between the surface of the protein chip and the protein molecule, and reduce the impact of the surface of the protein chip on the spatial conformation of the protein molecule; the biological activity of the protein immobilized by this method is higher than that obtained by physical The biological activity of the protein adsorbed or simply fixed by component B is more than doubled.

Description

technical field [0001] The invention relates to a method for immobilizing proteins on the surface of a protein chip. Background technique [0002] Protein is a kind of organic macromolecule with complex composition and structure, which can interact with and adsorb on almost all solid surfaces. Immobilizing target protein molecules on the surface of protein chips has been widely used in biomedical fields such as protein purification, solid-phase immunoassay and biomaterial preparation. [0003] At present, the main method of immobilizing proteins on the surface of protein chips is through physical adsorption, that is, through the interaction of electrostatic, hydrophobic and hydrogen bonds between the two to immobilize proteins on the solid surface. As described in Jonsson, U.; Ivarsson, B.; Lundstrom, I.; Berghem, L J. Colloid. Chlorodimethylsilane modified silicon surface. However, using the method of physical adsorption, the biological activity of protein molecules immo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/543C12Q1/68
Inventor 王战会靳刚
Owner INST OF MECHANICS CHINESE ACAD OF SCI
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