A kind of small molecule microarray and preparation method thereof

A technology of small molecules and microarrays, applied in the field of small molecule microarrays and its preparation, can solve the problems of detection signal reduction, electrostatic adsorption, and increasing sources of non-specific adsorption

Active Publication Date: 2015-08-19
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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  • A kind of small molecule microarray and preparation method thereof
  • A kind of small molecule microarray and preparation method thereof
  • A kind of small molecule microarray and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Preparation of Brush-like PEG Surface Small Molecule Microarray

[0075] Specific steps:

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

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

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

[0079] (4) 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 bipyridyl, 5 mM polyethylene glycol methacrylate (molecular weight 360), and methanol and water as solvents 1:1 (v / v) solvent mixture.

[0080] (5) Soak the gold flakes in the prepared ...

Embodiment 2

[0089] Embodiment 2 Preparation of hyperbranched PEG surface small molecule microarray

[0090] Specific steps are as follows:

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

[0092] (2) After the predetermined reaction time is reached, take out the glass and clean it.

[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 bipyridyl, 5 mM polyethylene glycol methacrylate (molecular weight 360), and methanol and water as solvents 1:1 (v / v) solvent mixture.

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

[0095] (5) After the predetermined growth time is reached, take out the gold sheet and rinse it with ethanol and water alterna...

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 substrate 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). Mix the two solutions at a volume ratio of 10:1, put the chip into the mixed solution, and incubate at 4°C for 12 hours. After reaching the predetermined time, take it out, wash it alternately with ethanol and deionized water, and dry it with nitrogen.

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

[0105] (4) Take out the chip, put it directly into the saturated α-cyclodextrin aqueous solution c...

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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 biochips, 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 developed rapidly in the past decade. Small molecule microarray refers to a high-density microarray formed by spotting (or printing) on ​​a solid surface and then immobilizing various organic small molecules—similar to the widely popular DNA microarray, a glass (or plastic) backing Microarrays of tens of thousands of different compounds can be printed on the substrate. Thanks to the miniaturization of analytical instruments, thousands of biochemical interactions can be simultaneously analyzed in one step. In 1999, Schreiber et al. reported the small molecule array for the first time, which was used as a probe and successfully applied to the detection of FKBP12 and its ligands (Koehler, A.N., A.F. Shamji, and S.L. Sc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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