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Small-molecule microarray and preparation method thereof

A small molecule and microarray technology, applied in the field of small molecule microarray and its preparation, can solve the problems of reducing detection signal, electrostatic adsorption, increasing the source of non-specific adsorption, etc.

Active Publication Date: 2014-05-14
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, the technical route of selective or directional immobilization of small molecule probes has fundamental defects: the active site of small molecule probes is likely to be fixed, thus losing the biochemical activity with the target, and may miss good active compounds
In such a three-step reaction, the final yield is the product of the yield of each step of the reaction, resulting in a substantial decrease in the final yield, thereby reducing the amount of immobilization of small molecules, which will lead to a decrease in the detection signal; at the same time, those not involved in the reaction The carboxylated end will make the surface have a certain amount of charge in the detection solution, which is easy to cause electrostatic adsorption and increase the source of non-specific adsorption

Method used

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  • Small-molecule microarray and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Preparation of brush-shaped PEG surface small molecule microarrays

[0075] Specific steps:

[0076] (1) A 3.5nm-thick chromium layer and a 50nm-thick gold layer were prepared on a glass substrate by thermal evaporation, as the substrate of the biochip.

[0077] (2) Prepare 400 μL of mono-mercapto-halothiol solution with a concentration of 1 mM as an initiator, prepare 400 μL of mono-mercapto-polyethylene glycol with the same concentration, mix the two solutions in a ratio of 1:99, and spread them on the surface of the chip. Incubate at °C for 12 hours.

[0078] (3) Wash the gold pieces alternately with ethanol and deionized water.

[0079] (4) Preparation of monomer solution: a total of 20 mL of monomer solution, of which ascorbic acid 0.04mM, copper chloride 0.04mM, bipyridine 0.8mM, polyethylene glycol methacrylate (molecular weight 360) 5mM, the solvent is methanol and water 1:1 (v / v) mixed solvent.

[0080] (5) Immerse the gold flakes in the prepare...

Embodiment 2

[0089] Example 2 Preparation of hyperbranched PEG surface small molecule microarrays

[0090] Specific steps are as follows:

[0091] (1) Prepare a glass substrate chip, clean and dry the chip and cover it with a certain amount of initiator solution.

[0092] (2) After reaching the predetermined reaction time, the glass is taken out and cleaned.

[0093] (3) Preparation of monomer solution: a total of 20 mL of monomer solution, including 0.04 mM ascorbic acid, 0.04 mM copper chloride, 0.8 mM bipyridine, 5 mM polyethylene glycol methacrylate (molecular weight 360), and the solvents are methanol and water 1:1 (v / v) mixed solvent.

[0094] (4) Immerse the gold flakes in the prepared monomer solution, and place them in an oxygen-free environment to grow polymers. The growth time is 6 hours, and the thickness of the polymer film is adjusted by controlling the growth time.

[0095] (5) After reaching the predetermined growth time, the gold pieces were taken out and rinsed alter...

Embodiment 3 3

[0101] Example 3 Three-dimensional polyrotaxane surface small molecule microarray

[0102] (1) As in step 1 in Example 1, a gold sheet was prepared as the base of the biochip.

[0103] (2) Prepare two ethanol solutions of mercaptopolyethylene glycol with a concentration of 1mM and different molecular weights, one is HS-(CH 2 ) 11 -EG6, the other is HS-EGn-COOH (molecular weight 2000). The two solutions were mixed at a volume ratio of 10:1, and then the chip was placed in the mixed solution and incubated at 4°C for 12 hours. After reaching the predetermined time, after taking out, alternately wash with ethanol and deionized water, and blow dry with nitrogen.

[0104] (3) The chip was immersed in an aqueous solution (20 mL) of 128 mM α-cyclodextrin and incubated for 4 hours, and then 750 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide was added to the solution. And N-hydroxysuccinimide 115mg, reaction 15min.

[0105] (4) Take out the chip and put it directly into a sa...

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Abstract

The invention discloses a preparation method for a small-molecule microarray. The preparation method comprises the steps of bonding the hydroxyl terminal of a nonspecific adsorption resistance material modified on a chip substrate with the hydroxyl terminal of a small-molecule light crosslinking agent through an esterification reaction, and randomly fixing small molecules on the hydroxyl terminals through light crosslinking. By the adoption of a direct esterification mode, the small-molecule light crosslinking agent is quickly and efficiently coupled to the surface of the nonspecific adsorption resistance material; a chip modification process is simple and convenient and can be implemented under a normal temperature condition; therefore, the modification cost is reduced, and the modification efficiency is improved; a high detection signal is guaranteed; a non-reacted group still keeps high nonspecific adsorption resistance capacity, so that a background signal is reduced; the prepared small-molecule microarray is high in detection signal and low in background signal and can be used for mutual action between high-flux screened small-molecule medicaments and targets.

Description

technical field [0001] The invention relates to the technical field of biological chips, in particular to a small molecule microarray and a preparation method thereof. Background technique [0002] Small molecule microarray is a high-throughput drug screening technology that has developed rapidly in the past decade. Small molecule microarrays refer to high-density microarrays formed by spotting (or printing) and then immobilizing various types of small organic molecules on a solid surface - similar to the widely popular DNA microarray, a glass (or plastic) lining Microarrays of tens of thousands of different compounds can be printed on the bottom, and thanks to the miniaturization of analytical instruments, thousands of biochemical interactions can be analyzed simultaneously in one step. In 1999, Schreiber et al first reported a small molecule array, which was used as a probe and successfully applied to the detection of FKBP12 and its ligands (Koehler, A.N., A.F.Shamji, and...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/531G01N33/68
CPCB01L3/502707B01L2200/0631B01L2300/168
Inventor 朱劲松杨墨程志强李少鹏周文菲
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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