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Supramolecular self-assembly biological chip, and preparation method and application thereof

A technology of supramolecular self-assembly and biochip, which is applied in biochemical equipment and methods, biological testing, microbial measurement/inspection, etc. It can solve the problems of poor performance of protein non-specific adsorption, difficulty in derivation, single spatial structure, etc. problems, to achieve the effect of controllable density of three-dimensional space structure, superior comprehensive performance, and wide selection space

Inactive Publication Date: 2012-07-04
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 dextran used in this method is a cross-linked network structure with a single spatial structure, which has a great influence on the immobilization form of biological signal molecules, the density and types of functional groups are greatly restricted, and the derivation is difficult; The performance of non-specific adsorption is not good; and there are many production steps, and the highly toxic epichlorohydrin is used in the production, the controllability, repeatability and safety of the chip are all limited, and the cost is expensive, resulting in excessive experimental costs. High, many units of Biacore instruments are idle

Method used

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  • Supramolecular self-assembly biological chip, and preparation method and application thereof
  • Supramolecular self-assembly biological chip, and preparation method and application thereof
  • Supramolecular self-assembly biological chip, and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0087] Choose metal-clad glass as the substrate, first vapor-deposit or sputter a 1-2nm chromium film on one surface of the glass substrate, and then vapor-deposit or sputter a 45-55nm thick gold film on the chromium film to prepare a bare For the gold chip, use an MV / Ozone cleaning machine to clean it with ozone ultraviolet rays to remove surface impurities, or wash it with ethanol and water several times, and then dry it with nitrogen gas and store it for later use.

[0088] Preparation of HS-PEG-OCH 3 (molecular weight=2000), HS-PEG-OH (ethylene glycol unit number is 6) and the solution that the molar ratio of the two is 10:1, 1:1, 1:10, the solvent can be water or PBS, and the total concentration is equal to It is 1mM, printed on the detectable area of ​​the bare gold chip by a chip printer, five spots are made for each solution, and a total of five lines are placed, incubated at 4°C for 1 hour, and then washed with ethanol and water to remove molecules that are not fixed ...

Embodiment 2

[0100] In this case, the substrates are glass slides, silicon wafers, quartz, polystyrene, and polydimethylsiloxane. The surface is cleaned with ethanol and water, and dried with nitrogen. The chip is prepared according to the following steps:

[0101] 1. Put it into 3-aminopropyltriethoxysilane (APES) diluted with 1:50 acetone, stay for 20-30 seconds, take it out and stop for a while, then put it into pure acetone solution to remove unbound APES, 3-aminopropyltriethoxysilane (APES) is bonded to the substrate to form a monomolecular layer on the surface of the substrate;

[0102] 2. Soak in 25% glutaraldehyde for 30 minutes, wash with acetone, immerse in 1 mM PEG (Mw=2000) with amino groups at both ends and PEG with amino group at one end and hydroxyl group at the other end (Mw=400) and both Incubate in an aqueous solution with a molecular ratio of 10:1 for 1 hour, then take it out and wash it with water to remove the unfixed diamino-PEG on the surface;

[0103] 3. Immerse it...

Embodiment 3

[0108] In this case, metal-coated glass is selected as the substrate; in this case, PEG containing dithiol is selected, and the specific structure is shown in the figure below:

[0109]

[0110] Wherein the first chain molecule PEG molecule R is -NH2, n=44; the second chain molecule PEG molecule R is -OH, n=5.

[0111] The chip preparation steps are as follows:

[0112] 1. First evaporate or sputter a 1-2nm chromium film on one surface of the glass substrate, then evaporate or sputter a 45-55nm thick gold film on the chromium film to prepare a bare gold chip, and use MV / Ozone The washing machine performs ozone ultraviolet cleaning to remove surface impurities, or washes with ethanol and water for many times, and then blows dry with nitrogen and saves it for later use;

[0113] 2. Prepare 5ml of α-cyclodextrin saturated aqueous solution, add PEG molecule (n=44) solid 100mg or a small amount of aqueous solution into the cyclodextrin solution, stir at room temperature for 2 h...

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Abstract

The invention provides a supramolecular self-assembly biological chip. The supramolecular self-assembly biological chip comprises a substrate (1), first chain molecules (3) which are fixed on the surface of the substrate, and second chain molecules (4) which are fixed on the surface of the substrate, wherein the first chain molecules and the second chain molecules are fixed on the substrate at intervals; part of one end, which is far away from the substrate, of each first chain molecule is included by cyclodextrin (2); the cyclodextrin comprises a functional group which can be directly or indirectly bound with protein molecules; and each second chain molecule is not longer than each first chain molecule. Poly(pseudo)rotaxane which is formed by coating the cyclodextrin and the first chain molecules and the second chain molecules which are arranged uniformly at the intervals are arranged on a self-assembly layer on the surface of the biological chip, a three-dimensional space has a dense, controllable and changeable structure, and compared with a two-dimensional surface, the three-dimensional space can fix more detection molecules such as the protein molecules. Moreover, the invention also provides a preparation method and application of the biological chip.

Description

technical field [0001] The invention relates to the field of biochips, in particular to a supramolecular self-assembled biochip, which can perform biochemical detection by combining with a surface plasmon resonance sensor. Background technique [0002] Biochip technology is a new technology developed by the intercrossing penetration of life science and microelectronics in recent years. With the continuous breakthrough of the Human Genome Project (HGP) research, this technology has been widely used in genetic diagnosis, functional gene research, genome library graphic analysis, disease diagnosis, proteomics research, immune detection, research and development of new drugs, Forensic science and many other fields. Biochip technology mainly establishes various arrayed gene or protein chips through surface chemical modification to achieve accurate, rapid, and large-information detection of cells, proteins, nucleic acids, and other biological components. It has high parallelism a...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/63C12Q1/68
Inventor 朱劲松王艳梅宋炉胜王坤程志强周大苏
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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