Method for fixing biometric identification molecules to biochip

A biochip and biorecognition technology, applied in chemical instruments and methods, analytical materials, laboratory containers, etc., can solve the problems of reduced activity of biorecognition molecules, reduction of bioanalysis technology performance, and hindrance of biomolecular affinity , to achieve the effect of avoiding non-specific adsorption, compact structure and uniform distribution

Active Publication Date: 2016-08-17
BEIJING RUILIANAN TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The physical adsorption method is the easiest way to immobilize biorecognition molecules such as DNA, antibody / antigen, etc. on the biochip. The disadvantages are that the immobilized biorecognition molecules have less molecular weight and are easy to fall off, and the non-directional adsorption of biorecognition molecules leads to loss of activity, etc.
The embedding method is a method of embedding biorecognition molecules with high-molecular organic polymers and immobilizing them on biochips. The disadvantage of this method is very obvious, that is, high-molecular organic polymers can hinder the affinity of biomolecules, and at the same time Large amount of non-specific adsorption, so it is less used in the preparation of biochips
The covalent bond method uses chemical bonding to immobilize biorecognition molecules such as antibodies / antigens, DNA, molecular imprinting molecules, and nucleic acid aptamers on biochips. It is characterized by firm binding and reusable biochips, but biorecognition molecules The activity will be reduced due to chemical bonding, and the non-directional immobilization of biorecognition molecules to the biochip can cause the loss of its activity, thereby reducing the performance of bioanalytical technology

Method used

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  • Method for fixing biometric identification molecules to biochip
  • Method for fixing biometric identification molecules to biochip
  • Method for fixing biometric identification molecules to biochip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 directly immobilizes the coated antigen on the surface of the biochip:

[0053] This example takes Microcystin-LR coated antigen (MC-LR-OVA) as an example, Microcystin-LR coated antigen was purchased from Sigma-al (China), according to figure 1 The schematic diagram of the immobilization process on the surface of the biochip is shown as follows:

[0054] 1) First use Piranha solution (concentrated H 2 SO 4 and H 2 o 2 The volume ratio is 3:1) at 25°C to clean the surface of the biochip (quartz glass sheets on the market, with a size of 1cm×1cm) for 30min to remove organic matter on the surface and to hydroxylate the surface; then Then put it into an ultrasonic cleaner and wash it thoroughly with ultrapure water until the pH value of the cleaning solution is neutral. Finally, dry it with nitrogen at room temperature, put it in a drying oven at 105°C for 2 hours, and then store it in Reserved in a vacuum drying oven;

[0055] 2) Put the clean surface hydro...

Embodiment 2

[0059] Embodiment 2 directly immobilizes the antibody on the surface of the biochip:

[0060] This example takes the bisphenol A antibody as an example. The bisphenol A antibody was purchased from Beijing Jinda Qingchuang Environmental Technology Co., Ltd., according to figure 1 The schematic diagram of the immobilization process on the surface of the biochip is shown as follows:

[0061] 1) First use Piranha solution (concentrated H 2 SO 4 and H 2 o 2 The volume ratio is 3:1) at 25°C to clean the surface of the biochip (quartz glass sheets on the market, with a size of 1cm×1cm) for 30min to remove organic matter on the surface and to hydroxylate the surface; then Then put it into an ultrasonic cleaner and wash it thoroughly with ultrapure water until the pH value of the cleaning solution is neutral. Finally, dry it with nitrogen at room temperature, put it in a drying oven at 105°C for 4 hours, and store it in Reserved in a vacuum drying oven;

[0062] 2) Put the clean ...

Embodiment 3

[0066] Example 3 DNA fragments are directly immobilized on the surface of the biochip:

[0067] The DNA fragment is: 5'-NH 2 -TTTTTTTTTT-3' (provided by Sangon Bioengineering (Shanghai) Co., Ltd.), according to figure 1 The schematic diagram of the immobilization process on the surface of the biochip is shown as follows:

[0068] 1) First use Piranha solution (concentrated H 2 SO 4 and H 2 o 2 The volume ratio is 3:1) at 25°C to clean the surface of the biochip (quartz glass sheets on the market, with a size of 1cm×1cm) for 30min to remove organic matter on the surface and to hydroxylate the surface; then Then put it into an ultrasonic cleaner and wash it thoroughly with ultrapure water until the pH value of the cleaning solution is neutral. Finally, dry it with nitrogen at room temperature, put it in a drying oven at 105°C for 3 hours, and store it in Reserved in a vacuum drying oven;

[0069] 2) Put the clean surface hydroxylated biochip into 2% dihydro-3-[3-(triethox...

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Abstract

The invention relates to a method for fixing biometric identification molecules to a biochip. The method comprises the following steps: (1) carrying out surface hydroxylation on the biochip; (2) carrying out silanization on the surface-hydroxylated biochip by using a silanization reagent containing a succinic anhydride group; (3) hydrolyzing the succinic anhydride group of the surface of the silanized biochip so as to form a carboxyl group; (4) activating the carboxylated biochip by using 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride and N-hydroxyl succinimide; and (5) connecting the biometric identification molecules to the surface of the activated biochip, thereby obtaining the biochip with the surface modified with the biometric identification molecules. According to the method, by adopting the silanization reagent containing the succinic anhydride group, the amino-containing biometric identification molecules can be conveniently and simply connected, so that the biometric identification molecules can be arranged in an oriented manner, a uniform and dense unimolecular structure is formed, and the identification efficiency of the biometric identification molecules is increased; and meanwhile, non-specific adsorption is avoided, so that the detection effect of the biochip is effectively improved.

Description

technical field [0001] The invention belongs to the field of biochip modification, and in particular relates to a method for immobilizing biorecognition molecules to biochips. Background technique [0002] Immobilizing biorecognition molecules efficiently on biochips is the key to developing high-sensitivity and high-specificity bioanalysis techniques. In recent years, bioanalysis technologies based on biochips and biosensors have been widely used in drug development, environmental monitoring, food testing, medical diagnosis and other fields due to their high sensitivity, strong specificity, and large detection throughput. A crucial step in most bioanalysis techniques is to immobilize specific biological recognition molecules (such as DNA, antibodies or antigens, nucleic acid aptamers, molecular imprinted molecules, etc.) Sensitive membrane of the target object. Methods for immobilizing biorecognition molecules on biochips include physical adsorption, embedding, and covale...

Claims

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Application Information

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IPC IPC(8): B01L3/00G01N33/543
CPCB01L3/502707B01L2200/10B01L2200/12G01N33/54353
Inventor 龙峰
Owner BEIJING RUILIANAN TECH CO LTD
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