Method for modifing protein chip surface

A protein chip, protein chip technology, applied in biochemical equipment and methods, microbial determination/inspection, material inspection, etc., can solve problems such as protein instability, low protein biological activity, protein spatial conformation change, etc. Guaranteed biological activity, reduced electrostatic interactions, improved properties

Active Publication Date: 2005-03-23
INST OF MECHANICS - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the defects that the existing methods for fixing proteins cause large changes in protein spatial conformation, low protein biological activity, and unstable proteins. In order to stably fix protein molecules on the surface and maintain their biological activity, and can inhibit the non-specific adsorption of protein molecules on solid surfaces, thus providing a method for modifying the surface of protein chips

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Fix small molecular weight protein molecules—insulin molecules with a molecular weight of 5,000 Daltons to the surface of the silicon substrate of the aldehyde-modified protein chip by covalent bonds, and reach saturation to obtain a protein chip with small molecular weight protein molecules immobilized ;

[0017] Soak the above-mentioned protein chip with immobilized insulin molecules in glutaraldehyde solution for 30 minutes to activate, and some amino groups of insulin molecules react with glutaraldehyde to make the surface of insulin molecules have aldehyde groups;

[0018] Prepare 1 mg / ml protein molecule to be immobilized——PBS (phosphate buffer) solution of human immunoglobulin G, which contains 0.05wt% Tween 20; put the activated protein chip immobilized with insulin molecule into the human immunoglobulin Soak in the globulin G solution for 30 minutes to immobilize human immunoglobulin G on the surface of the protein chip.

Embodiment 2

[0020] A small molecular weight protein molecule - bovine serum albumin molecule with a molecular weight of 50,000 Daltons is covalently fixed on the surface of the gold substrate of the aldehyde-modified protein chip, reaching saturation, and a small molecular weight protein molecule is immobilized protein chip;

[0021] Soak the above-mentioned protein chip with immobilized bovine serum albumin molecules in an ethylcarbodiimide (EDC) solution for activation for 10 minutes, and some carboxyl groups of insulin molecules react with ethylcarbodiimide to form active esters;

[0022] Prepare 1 mg / ml protein molecule to be immobilized——PBS (phosphate buffer) solution of human serum albumin, which contains 0.05wt% Tween 20; put the above-mentioned activated protein chip immobilized with bovine serum albumin molecule Soak in the serum protein solution for 30 minutes to immobilize human serum protein on the surface of the protein chip.

Embodiment 3

[0024] A small molecular weight protein molecule - a lysozyme molecule with a molecular weight of 30,000 Daltons, is covalently fixed on the surface of the plastic substrate of the aldehyde-modified protein chip to achieve saturation, and a protein with a small molecular weight protein molecule is immobilized chip;

[0025] Soak the protein chip on which the lysozyme molecule has been immobilized in an ethylcarbodiimide (EDC) solution for 10 minutes to activate, and some carboxyl groups of the lysozyme molecule react with ethylcarbodiimide to generate an active ester;

[0026] Prepare 1mg / ml of protein molecules to be immobilized—monoclonal antibody in PBS (phosphate buffer) solution, which contains 0.05wt% Tween 20; put the activated protein chip immobilized with lysozyme molecule into the monoclonal antibody Soak in the solution for 30 minutes to immobilize the monoclonal antibody to the surface of the protein chip.

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Abstract

The invention discloses a method to modify proteid chip surface, its character is fixing a layer of low molecular weight proteid molecule on the chip by normal method at first, then fixing the proteid chip which is needed to be fixed. The low molecular weight is 1X10 to the power 3 -9X10 to the power 5 Dalton. The method can modify the surface character of the proteid chip surface: on one hand, because of the covering of the low molecular weight molecule on the chip surface, the biology activity of the unfixed proteid molecule can be ensured, and at the same time, the aspecfic absorb is controlled on the surface; on the other hand, the low molecular weight molecule has amino, carboxyl, hydroxyl and other active group polar aminophenol on the proteid molecule surface, they can be used to fix porteid molecule directly after being activeted, and to form stable covalent bond.

Description

technical field [0001] The invention relates to a method for modifying the surface of a protein chip. Background technique [0002] With the development of science and technology, protein chips have been widely used in many biomedical fields. Commonly used protein chips are to immobilize target protein molecules on the surface of a solid substrate. Protein is a kind of organic macromolecule with complex composition and structure, which can interact with and adsorb on almost all solid surfaces. In the fabrication of protein chips, it is not only necessary to stably immobilize protein molecules on the surface to maintain their biological activity, but also to be able to effectively inhibit non-specific adsorption on the surface. [0003] At present, the methods of immobilizing proteins on solid surfaces are mainly through physical adsorption or covalent linkage. As described in Jonsson, U.; Ivarsson, B.; Lundstrom, I.; Berghem, L.J. Colloid. Interactions such as bonds secu...

Claims

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Application Information

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