Protein chip for covalent fixing biomolecular and its preparation method

A protein chip and biomolecule technology, applied in the field of protein chips for biological detection, can solve the problems of false positives, loss of biological activity, poor stability of ligand molecules, etc., to avoid false positive detection results, inhibit physical adsorption, and be less prone to denaturation loss. live effect

Inactive Publication Date: 2005-09-21
INST OF MECHANICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ligand molecules immobilized on the surface of the chip by physical adsorption method have poor stability, are easy to fall off, and easily lead to denaturation of ligand molecules and loss of biological activity.
In addition, non-target proteins are easily adsorbed on the surface of protein chips immobilized with ligand molecules by physical adsorption, resulting in false positive results

Method used

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  • Protein chip for covalent fixing biomolecular and its preparation method
  • Protein chip for covalent fixing biomolecular and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] according to figure 1 Preparation of a protein chip for detection of human immunoglobulin G (IgG).

[0032] The solid substrate 1 of the chip is a 0.5mm polished silicon wafer. The modified layer 2 is composed of chlorosilane polyethylene glycol derivatives, which are respectively Cl(CH 3 ) 2 Si(CH 2 ) 11 (OCH 2 CH 2 ) 3 OCH 2 COOCH 2 CH 3 and Cl(CH 3 ) 2 Si(CH 2 ) 11 (OCH 2 CH 2 ) 3 OCH 2 CH 3 , Made by mixing 1:50 (molar ratio). The ligand molecule is an antibody (antiIgG) to human immunoglobulin G (IgG).

[0033] The preparation of the protein chip is carried out as follows:

[0034] 1. Cleaning treatment of silicon wafer surface

[0035] Clean the surface of the silicon wafer with a weak alkaline solution (such as ammonia water) to remove surface stains.

[0036] 2. Hydrophilic treatment of silicon wafer surface

[0037] Put the silicon chip processed in step 1 into 30wt% H with a volume ratio of 1:3 2 o 2 and 98 wt% H 2 SO 4 soaked in th...

Embodiment 2

[0045] Preparation of protein chip for detection of human insulin (insulin).

[0046] The solid substrate of the chip is a 0.5mm polished silicon wafer, and the surface is a naturally generated silicon dioxide film. The derivatives of polyethylene glycol are respectively Cl 2 CH 3 Si(CH 2 ) 9 (OCH 2 CH 2 ) 4 OCH 2 COOCH 2 CH 3 and Cl 2 CH 3 Si(CH 2 ) 9 (OCH 2 CH 2 ) 3 OCH 3 . Ligand molecules are insulin antibodies (anti-insulin).

[0047] The preparation of the protein chip is carried out as follows:

[0048] 1. Cleaning treatment of silicon wafer surface

[0049] Clean the surface of the silicon wafer with a weak alkaline solution (such as ammonia water) to remove surface stains.

[0050] 2. Hydrophilic treatment of silicon wafer surface

[0051] Put the silicon chip processed in step 1 into 30wt% H with a volume ratio of 1:3 2 o 2 and 98 wt% H 2 SO 4 Soak in the mixed solution for 30 minutes, take it out and wash it with deionized water until neutr...

Embodiment 3

[0058] Preparation of protein chip for detection of interleukin 6 (IL-6). The solid substrate of the chip is a 0.5mm polished glass sheet. The derivatives of polyethylene glycol are respectively Cl 3 Si(CH 2 ) 12 (OCH 2 CH 2 ) 5 ONH 4 and Cl 3 Si(CH 2 ) 12 (OCH 2 CH 2 ) 3 OCH 3 . The ligand molecule is the receptor (IL-6R) for interleukin 6 (IL-6).

[0059] The preparation of the protein chip is carried out as follows:

[0060] 1. Cleaning treatment of glass surface

[0061] Clean the glass surface with a weak alkaline solution (such as ammonia) to remove surface stains.

[0062] 2. Hydrophilic treatment of glass surface

[0063] Put the glass treated in step 1 into 30wt% H with a volume ratio of 1:3 2 o 2 and 98 wt% H 2 SO 4 Soak in the mixed solution for 20 minutes, take it out and wash it with deionized water until neutral, and obtain a hydrophilic substrate with hydroxyl groups on the surface.

[0064] 3. Preparation of chlorosilane polyethylene glyc...

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Abstract

A protein chip with ligand moleculae fixed to the surface of solid substrate in covalent mode features that the chlorosilane polyethanediol derivative is used as the modifying layer to link the solid substrate in covalent mode, and the inducing ligand film is linked to the active groups of said chlorosilane polyethanediol derivative via covalent bonds. Its advantages are high stability of ligand moleculae, and high detection correctness.

Description

technical field [0001] The invention relates to a protein chip for biological detection, in particular to a protein chip capable of covalently immobilizing ligand molecules on a solid surface and effectively inhibiting physical adsorption of protein molecules on the chip surface and a preparation method thereof. Background technique [0002] Protein chip is a new technology to detect biomolecules by using ligand molecules immobilized on the chip surface. The mass of ligand molecules immobilized on the surface of the protein chip and the adsorption of other non-target proteins on the surface of the chip are important factors affecting the detection of the protein chip. At present, the methods used to immobilize ligand molecules are mostly physical adsorption methods, such as the immobilization methods used in detection methods such as ELISA and RIA. The ligand molecules immobilized on the surface of the chip by the physical adsorption method have po...

Claims

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Application Information

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