Preparation method of biosensor for detecting L-histidine and application thereof

A biosensor and histidine technology, applied in the field of analytical chemistry and biosensors, can solve the problems of needs, complex methods, and high detection costs, and achieve the effects of cheap price, simple operation, and high sensitivity

Active Publication Date: 2014-09-17
凯惠睿智生物科技(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These methods have their own advantages and can meet the detection requirements of L-histidine to varying d

Method used

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  • Preparation method of biosensor for detecting L-histidine and application thereof
  • Preparation method of biosensor for detecting L-histidine and application thereof
  • Preparation method of biosensor for detecting L-histidine and application thereof

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preparation example Construction

[0026] The present invention is achieved through the following measures: a preparation method of a biosensor for detecting L-histidine, comprising the following steps:

[0027] (1) Utilize existing method, prepare the colloidal gold of suitable particle size;

[0028] (2) Utilizing the effect of invertase and colloidal gold, the invertase is modified on the surface of colloidal gold;

[0029] (3) Utilize the effect of DNA2 and invertase-colloidal gold to modify the sulfhydryl-modified DNA2 onto invertase-colloidal gold;

[0030] (4) Using DNA1 and streptavidin-modified magnetic beads to modify DNA1-streptavidin on the magnetic beads to obtain DNA1-streptavidin-modified magnetic beads (DNA1-MBs);

[0031] (5) Using hybridization technology to construct biosensors.

[0032] The preparation of described DNA2-modified invertase-colloidal gold, DNA1-streptavidin-modified magnetic beads and sensors comprises the following steps:

[0033] (1) Activate 3.0 μmol of thiol-modified DNA2...

Embodiment

[0041] Embodiment: Biosensor detection L-histidine based on DNA hybridization technology

[0042] 1. Experimental part

[0043] 1.1 Instruments and reagents

[0044] 1.1.1 Instruments

[0045] Yicheng Blood Glucose Meter (Beijing Yicheng Bioelectronic Technology Co., Ltd.), Z-82A Air Bath Constant Temperature Oscillator (Quantan Medical Instrument Factory); Anke-TGL-16C Feiweng High-speed Centrifuge (Shanghai Anting Scientific Instruments) factory).

[0046] 1.1.2 Reagents

[0047] HAuCl 4 、Na 3 C 6 h 5 o 7 , TCEP (tris(2-carboxyethyl)phosphine hydrochloride), and sucrose were purchased from Sinopharm Chemical Reagent Co., Ltd.; invertase was purchased from Sigma; streptavidin-modified magnetic beads were purchased from Tianjin Beisi Le Chromatography Technology open your heart.

[0048] The DNA was synthesized by Saibaisheng Company, and the sequence is as follows:

[0049] DNA1:5'-GAG TGA TAT GAA GGA TGG GCG TGT CGG GGC TAT TCT CTA C-biotin-3'

[0050] DNA2:5'-GT...

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Abstract

The invention relates to a preparation method of a biosensor for detecting L-histidine and application thereof. Firstly, single strand DNA1 modified by biotin is fixed on magnetic beads (MBs) modified by streptavidin (SA) to obtain a compound DNA1-MBs. In addition, colloidal gold is modified by DNA2 and invertase to obtain DNA2-invertase modified colloidal gold. Then, DNA1-MBs and DNA2-invertase modified colloidal gold are mixed to obtain an L-histidine sensor. A certain amount of L-histidine is added into the prepared L-histidine sensor to react for a period of time, through magnetic separation, a supernatant is obtained, saccharose is added into the supernatant to react for a period of time and is converted to glucose by invertase, generated glucose is directly detected by a glucometer, and the content of L-histidine is detected by detecting the content of glucose. The sensor is high in sensitivity and selectivity; the glucometer is used as a detection instrument and the magnetic beads are used as carriers; the operation is simple; the detection instrument is small in size, low in price and convenient to carry.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry and biosensor, in particular to a preparation method and application of a biosensor for detecting L-histidine. In addition, the present invention also relates to the method for detecting L-histidine by the biosensor. Background technique [0002] L-Histidine (L-Histidine, L-His), as a neurotransmitter or neuromodulator, plays an important role in the central nervous system of mammals. Studies have shown that it controls the transmission and reduction of metals in important biological substrates. Internal bleeding from small wounds; excessive intake of L-histidine can lead to poisoning, and changes in the concentration of L-histidine in organisms can cause some diseases. Therefore, the detection of L-histidine will be very important important. [0003] In recent years, the L-histidine determination methods mainly include: electrochemical method (Junfei Liang, Zhengbo Chen, Lin Guo an...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/54346G01N33/6812
Inventor 混旭柏莉徐亚琼
Owner 凯惠睿智生物科技(上海)有限公司
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