Establishment method based on surface plasma reinforcing energy transferring biosensor

A surface plasmon and biosensor technology, which is applied in the field of biosensor construction, can solve the problems of biosensor limitations, complex surface functionalization chemical bonds and steps, etc., and achieve the effect of simple steps, overcoming the complex synthesis of nanomaterials, and stable fluorescence properties

Inactive Publication Date: 2014-08-06
NANKAI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Although the application of biosensors based on fluorescence resonance energy transfer is relatively mature, due to its unique effective distance of energy transfer (1.0-10.0 nm), complex surface functionalization and chemical bonds between donor and acceptor are required when designing energy transfer systems. and steps, leading to great limitations in the development of new and efficient biosensors

Method used

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  • Establishment method based on surface plasma reinforcing energy transferring biosensor
  • Establishment method based on surface plasma reinforcing energy transferring biosensor

Examples

Experimental program
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Effect test

Embodiment 1

[0025] A method for constructing a biosensor based on surface plasmon enhanced energy transfer, the steps are as follows:

[0026] 1) Preparation of protein-coated fluorescent gold nanoclusters:

[0027] Add 10 mL of 25.4 mmol / L chloroauric acid solution and 10 mL of 30 mg / mL trypsin solution in sequence in a 50 mL round bottom flask, and react in a 37°C water bath for 10 minutes with magnetic stirring in the dark, then add dropwise 1mL of sodium hydroxide solution with a concentration of 1mol / L, adjust the pH of the system to 12.0, and then continue to react for 36 hours to obtain the crude product of trypsin-coated fluorescent gold nanoclusters, and use the ultrafiltration membrane to filter the crude product at 6000 rpm Ultrafiltration and centrifugation at a speed of 1 / min for 30 minutes, discard the supernatant, redisperse the separated trypsin-coated fluorescent gold nanoclusters in ultrapure water, and store in the dark at 4°C;

[0028] 2) Preparation of mercaptoethyla...

Embodiment 2

[0033] A method for constructing a biosensor based on surface plasmon-enhanced energy transfer, the steps and methods are basically the same as in Example 1, the difference is that the protein is bovine serum albumin, and correspondingly prepared fluorescent gold nanoclusters coated with bovine serum albumin cluster.

Embodiment 3

[0035] A method for constructing a biosensor based on surface plasmon enhanced energy transfer, the steps and methods are basically the same as in Example 1, the difference is that the protein is lysozyme, and correspondingly prepared fluorescent gold nanoclusters coated with lysozyme.

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Abstract

The invention provides an establishment method based on a surface plasma reinforcing energy transferring biosensor. The establishment method comprises the following steps of: 1) preparing a fluorescent gold nanoparticle cluster wrapped with protein; 2) preparing the gold nanoparticle modified by mercaptoethylamine; and 3) establishing an experiment system based on the surface plasma reinforcing energy transferring biosensor. The establishment method based on the surface plasma reinforcing energy transferring biosensor provided by the invention has the advantages that the fluorescent gold nanoparticle cluster wrapped with the protein and the gold nanoparticle modified by mercaptoethylamine are introduced to be served as the donor and the receptor for energy transferring; the concept of surface plasma reinforcing energy transfer is utilized to achieve quick, accurate, high-sensitivity and high-selectivity quantifying of the object liquaemin; synthesizing of the nanometer material in the method is simple in steps, removes harsh equipment and condition, is safe, simple and convenient to operate, low in toxicity, and low in cost; and the used synthesizing and detecting instrument and equipment are common equipment; and a reaction condition is simple; and the reaction can be simply carried out at a room temperature of water bath at 37 DEG C.

Description

technical field [0001] The invention relates to the technical field of inorganic nanometer material preparation and spectrum application, in particular to a construction method of a biosensor based on surface plasmon enhanced energy transfer. Background technique [0002] Biosensor refers to a biochemical analysis device that is close to the molecular size and can give real-time signals when interacting with the analyte. Biosensors have the advantages of small size, low cost, no need for pretreatment, and no influence of external electromagnetic fields. High sensitivity and lower detection limit, better selectivity and less substance interference, higher accuracy and better precision, more complete and reliable speciation analysis, higher analysis speed and degree of automation , Miniaturized or non-destructive analysis with smaller sample volume, in situ, in vivo, real time and on line analysis, miniaturization, miniaturization and intelligence of analysis devices Wait. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 严秀平刘敬民
Owner NANKAI UNIV
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