Method for preparing acetylcholinesterase fluorescence biosensor

An acetylcholinesterase and biosensor technology is applied in the field of preparation of acetylcholinesterase fluorescent biosensors, which can solve the problems of inability to use the biosensor multiple times, high instrument cost, loss of enzymatic activity, etc., and achieves continuous and controllable operation and low cost. Inexpensive and simple process

Inactive Publication Date: 2009-11-18
王振宇
View PDF3 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems that the existing enzyme biosensors are affected by the environment, resulting in the loss of enzyme activity and false positives; the high price of enzymes, the inability to use the biosensor for multiple times due to irreversible reactions, and the high cost of instruments, etc., the present invention provides an acetylcholinesterase fluorescent The preparation method of the biosensor can effectively improve the speed and accuracy of the determination of organophosphorus pesticide residues in food

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0007] Specific embodiment 1: In this specific embodiment, 1. A preparation method of acetylcholinesterase fluorescent biosensor, characterized in that the steps of the preparation method of acetylcholinesterase fluorescent biosensor are as follows: 1. Acetylcholinesterase extraction: 1 , Store the purchased live crucian carp head in a self-made ice box, cut the skull, and then take out the entire brain tissue with tweezers, remove blood streaks with filter paper, and remove non-brain tissue, and place it in a watch glass for weighing. 2. Tris-HCl extraction method Homogenize under the condition of Tris-HCl ice bath containing 0.1% Triton X-100 and 0.05mol / L NaCl (every 15s, 5s interval). The homogenate was centrifuged (10000r / min) at 0~4℃ for 10min. 2. Purification: 1. Desalting. After ammonium sulfate fractionation and salting out, the enzyme solution dissolved in Tris-Hcl (0.07mol / l, pH=8) is put into a dialysis bag, and the distilled water is dialyzed for 48h, and BaCl is used...

specific Embodiment approach 2

[0008] Specific embodiment two: this specific embodiment is different from specific embodiment one in that it is characterized in that in step one, the extraction of acetylcholinesterase adopts pH=8.0~10.0Tris-HCl solution and homogenize 4~4 under ice bath conditions. 10min (every 15s, intermittent 5s), the homogenization ratio (mass:volume) is 1:5. Others are the same as the first embodiment.

specific Embodiment approach 3

[0009] Specific embodiment three: this specific embodiment is different from specific embodiment one in that it is characterized in that step two is purified by Sephadex G-200 gel chromatography column, and the eluent is 0.1-0.2 mol / L phosphate buffer , The flow rate is 0.15mL / min. Others are the same as the first embodiment.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a method for preparing an acetylcholinesterase fluorescence biosensor. The invention solves the following problems: the existing enzyme biosensor is prone to being affected by environmental influence so that the enzyme losses activity thus resulting false- positive; the existing biosensor is not allowed for repeated use due to irreversible reactions, in which enzymes are expensive, so that the cost of the equipment is excessively high; the selectivity is low, etc. The method comprises the following steps: firstly, carrying out the extraction separation and purification on acetylcholinesterase in crucian carp brain; then, carrying out the immobilization on the acetylcholinesterase in the crucian carp brain by chitosan; preparing immobilized acetylcholinesterase membranes, namely, obtaining the acetylcholinesterase fluorescence biosensor. The acetylcholinesterase fluorescence biosensor prepared by the invention dispenses with neither external power connection nor reference electrodes, and has the advantages of small size, light weight, high response speed, electromagnetic interference resistance, corrosion resistance, high sensitivity, wide measurement bandwidth, convenient operation, low cost and the like, therefore, the invention is applicable to the rapid detection of organophosphorus pesticide in food.

Description

Technical field [0001] The invention relates to a preparation method of an acetylcholinesterase fluorescent biosensor. Background technique [0002] my country is a large agricultural country, and pesticides are very important means of production in agricultural production. Because organophosphorus pesticides have the advantages of high efficacy, many varieties, many control objects, and easy degradation in the environment, they are widely used in my country's agricultural production. Organophosphorus pesticides and their residues often cause poisoning to humans and animals. To solve the key problem of agricultural products is the detection technology. The establishment of an accurate, fast, simple and cheap pesticide residue detection technology is a top priority for food safety and environmental protection. [0003] The development of rapid detection methods for organophosphorus biosensors has been reported at home and abroad. There are many developments of electrochemical enzy...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N27/327G01N33/48
Inventor 王振宇焦岩
Owner 王振宇
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap