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Method for fixing proteins on surface of solid

A solid surface, protein technology, applied to the immobilization of proteins, using aluminum nitride film to fix proteins, can solve the problems of high cost, less obvious immobilization effect, cumbersome steps, etc., and achieve low cost, easy long-term storage, high activity Good results

Active Publication Date: 2014-02-26
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above method also realizes the immobilization of proteins, there are also corresponding defects, mainly including: first, the steps are cumbersome; second, the immobilization effect is not obvious; third, the cost is relatively high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] This embodiment is implemented under the requirements of the following implementation conditions and technical conditions:

[0025] The solid substrate used in this embodiment is a silicon wafer covered with a 100 nm thick gold film.

[0026] The specific protein immobilization process is as follows:

[0027] (1) Put the silicon wafer into acetone, absolute ethanol, and deionized water in sequence, ultrasonically clean each for 3 minutes, take it out, and blow dry with nitrogen.

[0028] (2) A layer of aluminum nitride film with a thickness of 10 nanometers is deposited on the surface of the gold film by sputtering deposition technology.

[0029] (3) Add 0.2 mg / mL human IgG dropwise on the surface of the aluminum nitride film, and incubate at a constant temperature of 37° C. for 2 hours.

[0030] (4) Rinse three times with phosphate buffer solution with pH=7.0, and blow dry with nitrogen gas.

[0031] (5) Add 10 mg / mL bovine serum albumin blocking solution dropwise o...

Embodiment 2

[0037] This embodiment is implemented under the requirements of the following implementation conditions and technical conditions:

[0038] The solid substrate used in this example is a glass slide.

[0039] (1) Place glass slides in acetone, absolute ethanol, and deionized water in sequence, ultrasonically clean them for 3 minutes, take them out, and dry them with nitrogen.

[0040] (2) A layer of aluminum nitride film with a thickness of 50 nanometers is deposited on the surface of the gold film by sputtering deposition technology.

[0041] (3) Add 0.2 mg / mL human IgG dropwise on the surface of the aluminum nitride film, and incubate at a constant temperature of 37° C. for 2 hours.

[0042] (4) Rinse three times with phosphate buffer solution with pH=7.0, and blow dry with nitrogen gas.

[0043] (5) Add 10 mg / mL bovine serum albumin blocking solution dropwise on the aluminum nitride surface, and incubate at 37° C. for 1 hour.

[0044] (6) Rinse three times with phosphate b...

Embodiment 3

[0049] This embodiment is implemented under the requirements of the following implementation conditions and technical conditions:

[0050] The solid substrate used in this embodiment is a silicon wafer.

[0051] (1) Put the silicon wafer into acetone, absolute ethanol, and deionized water in sequence, ultrasonically clean each for 3 minutes, take it out, and blow dry with nitrogen.

[0052] (2) Patterning of AlN film:

[0053] Spin-coat photoresist 5 microns and perform photolithography to form a photoresist pattern opposite to the required aluminum nitride pattern.

[0054] A layer of aluminum nitride film with a thickness of 100 nm is deposited on the surface of the photoresist by sputtering deposition technology.

[0055] Soak the sample in acetone for 10 minutes to remove the photoresist and finally get the aluminum nitride pattern.

[0056] (3) Add 0.2 mg / mL human IgG dropwise on the surface of the aluminum nitride film, and incubate at a constant temperature of 37° C....

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Abstract

The invention discloses a method for fixing proteins on the surface of a solid, which relates to the field of a biosensor. The method comprises the following steps of: (1) cleaning the surface of the solid; (2) depositing an aluminium nitride thin film on the surface of the solid; (3) fixing the proteins, i.e. directly dropwise adding the proteins on the surface of the aluminium nitride thin film and washing after incubating. The method can be used for fixing the proteins on a biological chip and a biological probe in the biological field and has the advantages that the operation steps are simple, the cost is low, the proteins have high fixation quantity and high activity and are uniform and firm, and the graphical fixation can be realized.

Description

technical field [0001] The invention relates to a protein immobilization method, in particular to a method for immobilizing protein by using an aluminum nitride film, and belongs to the field of biological detection and sensors. Background technique [0002] In recent years, with the continuous development of biotechnology, the research and application of biosensors and biochips have become increasingly widespread. How to effectively immobilize proteins on biosensors and biochips is a key issue in biological detection. At present, the commonly used immobilization methods for immobilizing proteins on solid surfaces mainly include adsorption method, embedding method, covalent binding method, etc., such as (1) Hoyer-Hansen G, Hamers M J, Pedersen A N, etc. Loss of ELISA specificity due to biotinylation of monoclonal antibodies. Immunol Methods 2000, 235(1-2): 91-9; (2) Haodan, Wayne M.Mullett and JanuszPawliszyn, Biological sample anysis with immunoaffinity solid-phase microex...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K17/14C07K17/02
Inventor 徐东韩程章陈达徐航张亚非
Owner SHANGHAI JIAOTONG UNIV