Micro-array biochip, and preparation method and application of micro-array biochip

A biochip and microarray technology, applied in biological testing, material inspection products, measuring devices, etc., can solve problems such as the successful detection of small molecule drugs, and achieve the effect of efficient preparation

Inactive Publication Date: 2015-09-09
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, none of the above methods for chip chemical modification has successfully detected small molecule drugs on protein arrays

Method used

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  • Micro-array biochip, and preparation method and application of micro-array biochip
  • Micro-array biochip, and preparation method and application of micro-array biochip
  • Micro-array biochip, and preparation method and application of micro-array biochip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1 detects biotin (Biotin) on streptavidin (SA) microarray

[0069] 1. Modification of biochip substrate:

[0070] (1) prepare one deck of 3nm thick chromium layer and one deck of 47nm thick gold layer with the method of thermal evaporation on glass substrate;

[0071] (2) Prepare the ethanol solution HS-(CH 2 )11 -EG 6 -OH at a concentration of 1 mM;

[0072] (3) Clean the biochip substrate with ethanol or deionized water, and then put the biochip into a plasma cleaner for cleaning for 3 minutes;

[0073] (4) Soak the biochip in the prepared thiol solution and incubate at 4°C for 12 hours. After the predetermined time is reached, take out the biochip, wash it alternately with ethanol and deionized water, and dry it with nitrogen;

[0074] (5) Prepare 20 mL of the solution required for esterification and linking photocrosslinking agent, 10 mM carboxyl-terminal photocrosslinking agent, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) 10 mM, 4 -Dimethyla...

Embodiment 2

[0091] Example 2 Detection of Tacrolimus (FK506) on FKBP12 Protein Microarray

[0092] 1. Modification of biochip substrate

[0093] (1) Prepare a 3nm thick chromium layer and a 47nm thick gold layer on the glass substrate by thermal evaporation, as the substrate of the biochip;

[0094] (2) Prepare the ethanol solution HS-(CH 2 ) 11 -EG 6 -OH and HS-(CH 2 ) 11 -EG 6 -COOH, the concentration is 1mM, OH terminal thiol and COOH terminal thiol are mixed at a volume ratio of 999:1;

[0095] (3) Clean the biochip substrate with ethanol or deionized water, and then put the biochip into a plasma cleaner for cleaning for 3 minutes;

[0096] (4) Soak the biochip in the prepared thiol solution and incubate at 4°C for 12 hours. After the predetermined time is reached, take out the biochip, wash it alternately with ethanol and deionized water, and dry it with nitrogen;

[0097] (5) Prepare 20mL of surface activation aqueous solution, mix EDC (0.4M) and NHS (0.1M) according to the ...

Embodiment 3

[0115] Example 3 Detection of FK506 on FKBP12 and its mutant protein microarray

[0116] 1. Modification of biochip substrate

[0117] With embodiment 2.

[0118] 2. Preparation of protein microarray

[0119](1) Prepare the phosphate buffer solution of FKBP12 (wild type) protein with pH = 7.4 and the concentration of 0.5 mg / mL, and the phosphate buffer solution of 5 single amino acid mutants (M1-M5) with the same pH and concentration, and set aside ;

[0120] (2) Dip the prepared chip substrate into a mixed solution of EDC (0.4M) and NHS (0.1M) (1:1, v / v) to activate for 15 minutes, take out the chip, wash it quickly with deionized water, and dry it with nitrogen ;

[0121] (3) Put the chip and the six protein buffer solutions prepared in step (1) into the corresponding positions of the printer, adjust the parameters, and print. The ambient humidity is 70%, the needle diameter is 300 μm, the contact time is 1s, and the array is 6×12 (like image 3 a);

[0122] (4) Incub...

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Abstract

The invention relates to a micro-array biochip, and a preparation method and an application of the micro-array biochip. The micro-array biochip comprises a glass chip substrate and a modification layer. The modification layer comprises a chromium layer, a silver and / or gold layer, a self-assembly monomolecular layer, and a photo-cross-linking agent and high-molecular layer, which are overlaid in sequential on the surface of the glass chip substrate. The invention also provides a protein micro-array, a method for preparing the protein micro-array, and an application of the protein micro-array to detect micromolecule medicine. According to the invention, three-dimensional high-molecular surface modification of the micro-array biochip is completed by the method, and the fixation amount of protein on the surface of the micro-array biochip can be effectively improved, so that micromolecule, hard to detect in the prior art, can be detected without marking, in accuracy, and in high flux. The micro-array biochip is of great significance to the field, such as molecular biology study and medicinal development.

Description

technical field [0001] The invention relates to the technical field of biochips, in particular to a microarray biochip and its preparation method and application. Background technique [0002] Studying the interaction between small molecule drugs and protein targets is of great significance to basic scientific research and drug development. However, in practical applications, there is a situation where a drug targets multi-target interactions, such as the study of drug complexity and side effects, the optimization of lead compounds, and the new use of old drugs, etc. Therefore, the development of an efficient small molecule-target interaction detection method will play a role in promoting target activity evaluation and drug screening. [0003] At the level of molecular biology, drug development often requires the collection and evaluation of interaction information (such as kinetics and affinity) between lead compounds / drugs and protein targets. In recent years, surface pl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/552G01N33/68
Inventor 杨墨朱劲松李少鹏周文菲王瑞
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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