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Method for marking and detecting glucide biochip

A technology of sugar substances and biochips, which is used in biological testing, material inspection products, scattering characteristics measurement, etc. to achieve rapid stability, good selectivity, and good biocompatibility.

Inactive Publication Date: 2009-07-29
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But at present, the carbohydrate biochip mainly uses fluorescence as the detection method, so it still has certain shortcomings in terms of detection sensitivity and selectivity; expanding the detection method of the carbohydrate biochip is of great significance to its development (Nature, 2007, 4, 437-444)

Method used

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  • Method for marking and detecting glucide biochip
  • Method for marking and detecting glucide biochip
  • Method for marking and detecting glucide biochip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Labeling and detection of 4-aminophenyl-α-D-mannopyranoside (Man-α)

[0027] (1) Synthesis of peptide-modified gold nanoparticles

[0028] Cystylylalanylleucylaspartylaspartate and cystylalanylleucylaspartylaspartylglycyllysyl (biotinyl)glycine in molar ratio 9 : 1 prepares the mixed solution of polypeptide mixture, mixes the mixture of gold nanoparticles modified by sodium citrate and polypeptide mixed solution with a molar ratio of 1: 50000, and reacts at room temperature after 1-2 hours. ) to purify the reaction product by centrifugation to obtain polypeptide-modified gold nanoparticles; disperse the polypeptide-modified gold nanoparticles in a neutral phosphate buffer solution and store at 4°C;

[0029] (2) Making carbohydrate biochips

[0030] A 100μM-50mM 4-aminophenyl-α-D-mannopyranoside (Man-α) monosaccharide chip was produced on the SmartArrayer 48 chip production system provided by Beijing Boao Biotechnology Co., Ltd., through 1% (w / v ) bovine se...

Embodiment 2

[0038] Example 2: Labeling and detection of 4-aminophenyl-β-D-galactopyranoside (Gal-β)

[0039] (1) Synthesis of peptide-modified gold nanoparticles

[0040] Cystylylalanylleucylaspartylaspartate and cystylalanylleucylaspartylaspartylglycyllysyl (biotinyl)glycine in molar ratio 9 : 1 prepares the mixed solution of polypeptide mixture, mixes the mixture of gold nanoparticles modified by sodium citrate and polypeptide mixed solution with a molar ratio of 1: 50000, and reacts at room temperature after 1-2 hours. ) to purify the reaction product by centrifugation to obtain polypeptide-modified gold nanoparticles; disperse the polypeptide-modified gold nanoparticles in a neutral phosphate buffer solution and store at 4°C;

[0041] (2) Making carbohydrate biochips

[0042] 8 μ M-100 mM 4-aminophenyl-β-D-galactopyranoside (Gal-β) monosaccharide chip was produced on the SmartArrayer 48 chip production system provided by Beijing Boao Biotechnology Co., Ltd., passed 1% (w / v) Bovine...

Embodiment 3

[0050] Embodiment 3: labeling and detection of ribonuclease B (RNase B)

[0051] (1) Synthesis of peptide-modified gold nanoparticles

[0052] Cystylylalanylleucylaspartylaspartate and cystylalanylleucylaspartylaspartylglycyllysyl (biotinyl)glycine in molar ratio 9 : 1 prepares the mixed solution of polypeptide mixture, mixes the mixture of gold nanoparticles modified by sodium citrate and polypeptide mixed solution with a molar ratio of 1: 50000, and reacts at room temperature after 1-2 hours. ) to purify the reaction product by centrifugation to obtain polypeptide-modified gold nanoparticles; disperse the polypeptide-modified gold nanoparticles in a neutral phosphate buffer solution and store at 4°C;

[0053] (2) Making carbohydrate biochips

[0054] A glycoprotein chip of 0.16 μg / ml-1mg / ml ribonuclease B (RNase B) was produced on the SmartArrayer 48 chip production system provided by Beijing Boao Biotechnology Co., Ltd., passed through 1% (w / v) bovine serum albumin ( BSA...

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Abstract

The invention provides a saccharide biochip marking and detecting method. The method takes the polypeptide-modified gold nanoparticles as the marker of a saccharide biochip, takes resonance light scattering spectroscopy as the detecting means of the saccharide biochip, and marks the reaction process by the reaction of avidin and biotin. The method is versatile and characterized by high sensitivity being up to 32ng / mL, good selectivity and less sample consumption amount. The detection limit of monosaccharides obtained by the method is 8 mu M; the detection limit of agglutinin is 100pg / mL; the linear range of the monosaccharides and the agglutinin is 3 order of magnitudes; the detection limit of obtained glycoprotein is 32ng / ml; the detection limit of agglutinin is 25.6pg / mL; and the linear range of the glycoprotein and the agglutinin is 3 order of magnitudes. The method can realize the research among the monosaccharides, the glycoprotein detection, the saccharide and corresponding agglutinin.

Description

technical field [0001] The invention relates to a method for labeling and detecting carbohydrate biochips. technical background [0002] Due to the special physical and chemical properties of inorganic nanoparticles (gold, silver, quantum dots, etc.), they have been widely used in various fields of nanotechnology, bionanotechnology and biomedical detection technology in the past decade (Chemical Review, Chem.Rev ., 2004, 104, 293-346; Chem. Rev., 2005, 105, 1547-1562). Resonance light scattering (RLS) is a new instrumental analysis technique developed rapidly in recent years. It has attracted much attention because of its high sensitivity and simple operation (Science, Science, 1995, 269, 935-939; Science, Science, 2000, 289, 1757-1760). Biochip is currently an important tool for genomics and proteomics research, and has the advantages of high throughput and less consumption of test samples (Genome Biol. 2001, 2, research 0004.1-0004.13). But at present, the carbohydrate ...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N21/49
Inventor 王振新高晶清刘殿骏
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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