Production method of surface plasma resonance chip

A surface plasmon and chip technology, applied in the direction of material inspection products, phase influence characteristic measurement, scattering characteristic measurement, etc. Eliminate problems such as shedding, and achieve the effects of low preparation cost, reduced experimental cost, and uniform matrix

Inactive Publication Date: 2009-07-08
INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the large size of protein molecules, there are inevitably exposed areas on the surface of the gold film; and pre-crosslinking will affect the flatness of the chip surface matrix; and the strength of direct coupling is not high enough, protein molecules are prone to breakout Falling off; these will affect the use effect, stability and repeatability of the experimental results of the chip
In addition to the above methods, there are no reports of other methods for preparing surface plasmon resonance chips.

Method used

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  • Production method of surface plasma resonance chip
  • Production method of surface plasma resonance chip
  • Production method of surface plasma resonance chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] (1) Pretreatment of biological protein:

[0026] Add tris(2-formylethyl)phosphine hydrochloride to the bovine serum albumin solution, so that the final concentrations of bovine serum albumin and tris(2-formylethyl)phosphine hydrochloride in the mixed solution are respectively 0.5 μM and 100μM, react for 30 minutes to reduce the disulfide bond in the bovine serum albumin molecule;

[0027] (2) Carry out the construction of chip surface matrix:

[0028] a) putting the surface plasmon resonance sensor chip with a pure gold film on its surface into the above-mentioned pretreated bovine serum albumin solution and soaking for 60 minutes to obtain a chip with bovine serum albumin molecules;

[0029] b) Soak the above-obtained chip with bovine serum albumin molecules in a mixed aqueous solution containing 0.4M N-hydroxysuccinimide and 0.5M ethyl-dimethylaminopropyl-carbodiimide for 12 Minutes for surface crosslinking;

[0030] c) Soak the surface-crosslinked chip in an aqueo...

Embodiment 2

[0036] (1) Pretreatment of biological protein:

[0037] Add 2-mercaptoethanol to the ovalbumin solution, so that the final concentrations of ovalbumin and 2-mercaptoethanol in the mixed solution are 20 μM and 2 mM respectively, and react for 60 minutes to reduce the disulfide bond in the ovalbumin molecule;

[0038] (2) Carry out the construction of chip surface matrix:

[0039] a) putting the surface plasmon resonance sensor chip with a pure gold film on the surface into the above-mentioned pretreated ovalbumin solution and soaking for 40 minutes to obtain a chip with ovalbumin molecules;

[0040] b) Soak the above-obtained chip with ovalbumin molecules in a mixed aqueous solution containing 0.01M N-hydroxysuccinimide and 0.01M ethyl-dimethylaminopropyl-carbodiimide for 30 minutes carry out surface cross-linking;

[0041]c) Soak the surface-crosslinked chip in an aqueous solution containing 0.01M bromoacetic acid and 0.01M potassium hydroxide for 300 minutes to carboxymethy...

Embodiment 3

[0043] (1) Pretreatment of biological protein:

[0044] Add 2-mercaptoacetic acid and dithiothreitol to the bovine hemoglobin solution, so that the final concentrations of bovine hemoglobin, 2-mercaptoacetic acid and dithiothreitol in the mixed solution are 1mM, 0.8M and 0.5M respectively, and react for 15 minutes , so that the disulfide bond in the bovine hemoglobin molecule is reduced;

[0045] (2) Carry out the construction of chip surface matrix:

[0046] a) putting the surface plasmon resonance sensing chip with a pure gold film on the surface into the bovine hemoglobin solution pretreated above and soaking for 5 minutes to obtain a chip connected with bovine hemoglobin molecules;

[0047] b) Soak the above-obtained chip with bovine hemoglobin molecules in a mixed aqueous solution containing 2M N-hydroxysuccinimide and 1M ethyl-dimethylaminopropyl-carbodiimide for 8 minutes for surface interaction. couplet;

[0048] c) Soak the surface-crosslinked chip in an aqueous so...

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Abstract

The invention discloses a preparation method of surface plasma resonance chips. The preparation method is characterized in that a disulfide bond reducing agent is adopted to modify biological protein molecules containing hydrosulfide group and / or disulfide bond; then, protein substrate is established on the surface of a chip by means of metal surface molecule self-assembly technology; and further modification is carried out to realize the preparation of a novel sensing chip used in SPR biosensor. The preparation method has the advantages of low preparation cost, simple operation, less time consumption and safety and harmlessness; moreover, the prepared chip has the functions of commercialized CM5 chips, thereby providing new possibilities for solving the prior problems of fewer types of surface plasma resonance chips and high cost.

Description

technical field [0001] The invention belongs to the technical field of surface plasmon resonance sensing, in particular to a preparation method of a surface plasmon resonance chip. Background technique [0002] Biosensors based on surface plasmon resonance (SPR) technology have the advantages of real-time monitoring of biomolecular interactions, label-free, fast analysis, high sensitivity, simple pretreatment, and less sample consumption. It is widely used in the fields of proteomics, drug development, clinical diagnosis, food safety and environmental monitoring, and shows broad application prospects. Among the numerous SPR biosensors, the Swedish BIACORE series instruments occupy the largest market share, and correspondingly, their sensor chips are most widely used. At present, the most commonly used chip is a long-chain carboxylated dextran matrix (CM5) chip, which is suitable for coupling molecules with amino groups, sulfhydryl groups, aldehyde groups, hydroxyl groups or...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/48G01N21/55G01N21/41
Inventor 宋存先欧惠超罗昭锋周宏敏姜浩江海峰
Owner INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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