Preparation method of three-dimensional biological chip substrate

A biochip and substrate technology, applied in the field of preparation of biochip substrates, can solve problems such as the inability to immobilize biomolecules at high density and restrict the wide application of biochips, and achieve high probe immobilization efficiency, high immobilization density, and detection The effect of high sensitivity

Active Publication Date: 2015-07-08
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this type of surface has the disadvantage of not being able to immobilize biomolecules at a high density, and there are still huge challenges in terms of sensitivity and repeatability, which also restricts the wide application of biochips in clinical routine diagnosis.

Method used

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  • Preparation method of three-dimensional biological chip substrate
  • Preparation method of three-dimensional biological chip substrate
  • Preparation method of three-dimensional biological chip substrate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Preparation of three-dimensional structured aminated glass substrate

[0029] (1) Hydroxylation of the film base: put the glass slide in a mixed solution of KOH, water, and isopropanol (mass ratio 1:18:21), sonicate for 15 minutes, wash twice with deionized water, and then Rinse with acetone for later use.

[0030] (2) Silanization of sheet base: prepare the ethanol solution of vinyltrimethoxysilane and glacial acetic acid, wherein the mass concentration of vinyltrimethoxysilane is 1%, and the mass concentration of glacial acetic acid is 0.1%. The substrate was put into the above solution, soaked for 30min, then washed with ethanol and dried.

[0031] (3) Preparation of the three-dimensional structure of the film base: configure a mixed solution of a photoinitiator (1-hydroxycyclohexyl phenyl ketone), PEGDA and GMA, acetone is a solvent, and the mass concentration of the photoinitiator is 5%, PEGDA: The molar ratio of GMA is 10:1, and the mass fraction of the mixed so...

Embodiment 2

[0034] Preparation of Three-Dimensional Structured Aminated Cycloolefin Copolymer (COC) Substrates

[0035] (1) Hydroxylation of sheet base: Wash the COC film in ethanol and dry it in the air, sandwich a 30wt% ammonium persulfate aqueous solution between the biaxially stretched BOPP and the COC film to form a liquid layer of about 0.5 μm. Then irradiate the surface of BOPP with UV light for 2 min at room temperature, rinse the obtained modified COC membrane with deionized water, immerse in ultrapure water for 16 h, rinse with ultrapure water and dry at room temperature.

[0036] (2) Silanization of sheet base: prepare the ethanol solution of vinyltrimethoxysilane and glacial acetic acid, wherein the mass concentration of vinyltrimethoxysilane is 3%, and the mass concentration of glacial acetic acid is 0.05%, the surface activated The substrate was put into the above solution, soaked for 25min, then washed with ethanol and dried.

[0037] (3) Preparation of the three-dimension...

Embodiment 3

[0040] Preparation of three-dimensional structured aminated polymethylmethacrylate (PMMA) substrate

[0041](1) Hydroxylation of the sheet base: the PMMA film is cleaned in ethanol and dried, and a 30 wt % ammonium persulfate aqueous solution is sandwiched between the biaxially stretched BOPP and the PMMA film to form a liquid layer of about 0.5 μm. Then irradiate the surface of BOPP with ultraviolet light for 3 minutes at room temperature, rinse the obtained modified PMMA membrane with deionized water, and immerse in ultrapure water for 16 hours, rinse with ultrapure water and dry at room temperature.

[0042] (2) Silanization of sheet base: prepare the ethanol solution of vinyltrimethoxysilane and glacial acetic acid, wherein the mass concentration of vinyltrimethoxysilane is 5%, and the mass concentration of glacial acetic acid is 0.08%, the surface activated The substrate was put into the above solution, soaked for 20min, then washed with ethanol and dried.

[0043] (3) P...

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Abstract

The invention discloses a preparation method of a three-dimensional biological chip substrate, wherein the biological chip substrate can be made from a glass substrate or a polymer substrate. The preparation method includes following steps: (1) introducing hydroxyl groups onto the surface of the substrate and performing silanization to introduce a double bond; (2) performing ultraviolet light grafting onto the substrate of the double bond-introduced substrate to obtain the substrate having a certain thickness, wherein the surface of the substrate is provided with epoxy groups; (3) adding the treated substrate into a solution containing amino groups to perform a reaction to prepare the substrate of which the surface contains the amino groups. By means of the method, a three-dimensional amino substrate probe is high in fixation amount, is stable in combination, can overcome the defects that a conventional surface modification-silane coupling agent method for preparing a two-dimensional substrate is less in fixation amount, is high in detection limit and is instable in result. The method has a good application prospect in the field of biological chips.

Description

technical field [0001] The invention relates to a method for preparing a biochip substrate with a three-dimensional structure. Background technique [0002] Biochip technology has the characteristics of high throughput, high integration, miniaturization, parallelization, diversification and automation. It has been widely concerned in the fields of biology and medicine since it was reported. important application prospects. The chemical treatment of the substrate surface is the basis of chip preparation, and the appropriate surface chemical modification strategy is also the most direct way to achieve high-sensitivity chip preparation. At present, the commonly used carriers for biochips are mainly divided into inorganic (silicon wafers, glass slides, metals, etc.) A variety of monolayer two-dimensional surface functional groups such as aldehyde groups, amino groups, epoxy groups, and sulfhydryl groups are introduced to immobilize DNA / protein molecules. However, this type of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J7/12C08J7/18C03C23/00C08L69/00C08L25/06C08L33/12C08L63/00C08L45/00C08L83/04
Inventor 杨万泰马育红赵长稳冯清
Owner BEIJING UNIV OF CHEM TECH
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