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Preparation method of surface plasma resonance (SPR) biosensor chip

A biosensing and chip technology, applied in measuring devices, material analysis through optical means, instruments, etc., can solve problems such as the difficulty in detecting trace small molecular substances, restricting the wide application of SPR technology, and increasing the cost of instrument use. Excellent response efficiency and specificity, easy to promote, and fast detection of samples

Active Publication Date: 2015-02-18
BEIJING ZHONGLONGYICHENG TECH CO LTD
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  • Summary
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the SPR sensor chip used in the market is expensive, the production process is cumbersome, and it is difficult to detect trace small molecular substances, which virtually increases the cost of the instrument, reduces the utilization efficiency of the instrument, and greatly restricts the wide application of SPR technology.

Method used

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  • Preparation method of surface plasma resonance (SPR) biosensor chip
  • Preparation method of surface plasma resonance (SPR) biosensor chip
  • Preparation method of surface plasma resonance (SPR) biosensor chip

Examples

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Embodiment 1

[0025] see figure 1 , in the embodiment of the present invention, a kind of SPR biosensing chip, adopts the multi-layer self-assembly method to make, the chip hierarchical structure is the antigen 1 of the target analyte, bovine serum albumin 2 or antibody 7, nano gold in sequence from top to bottom 3. L-cysteine ​​4, nano gold 3, L-cysteine ​​4, gold film 5, glass matrix 6. Wherein the diameter of nano gold is 15-50nm.

[0026] The preparation method of described SPR biosensor chip, concrete steps are:

[0027] 1) self-assemble a layer of L-cysteine ​​4 on the surface of the glass matrix 6 coated with gold film 5, then connect the nano-gold 3 to the amino group of L-cysteine ​​4, repeat the above steps;

[0028] 2) Finally, the antigen 1 coupled to the antibody 7 or bovine serum albumin 2 is connected to the gold nanoparticles 3;

[0029] 3) The exposed gold nanoparticles 3 are blocked with bovine serum albumin 2 prepared in phosphate buffer saline, to obtain.

[0030] Th...

Embodiment 2

[0033] Adopt the SPR biosensor chip prepared by the present invention to detect the operation method of melamine content in milk, concrete steps are as follows:

[0034] (A) Sample processing method:

[0035] (A1) Take 50 μL of milk in a 2 mL centrifuge tube, add 950 μL of absolute ethanol, vortex and mix for 30 seconds, let stand for 2 minutes, and take the supernatant for the next step;

[0036] (A2) Take 100 μL of the above supernatant in a 1 mL centrifuge tube, dry it at 70°C or air-dry it naturally, add 100 μL of phosphate buffer (pH=7.4) at 70°C and fully dissolve it before testing.

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Abstract

The invention discloses a preparation method of a surface plasma resonance (SPR) biosensor chip. L-cysteine, nanogold and bovine serum albumin coupled antigen molecules are sequentially assembled on a substrate glass sheet plated with a gold membrane with the thickness of 50nm, so that the SPR biosensor chip is obtained. The preparation method of the SPR biosensor chip is simple, rapid and low in cost, no fussy activating treatment needs to be carried out, the SPR biosensor chip is applicable to detection of multiple trace small molecular substances of cyanurotriamide, clenbuterol hydrochloride, ampicillin and the like, the regenerability is good, and the SPR biosensor chip can be widely applied to the fields of food safety, environments, biological science and the like.

Description

technical field [0001] The invention relates to the field of optical sensor chip preparation, in particular to a preparation method of a surface plasmon resonance SPR biosensor chip. Background technique [0002] Surface plasmon resonance (SPR), a physical optics phenomenon, has been developed into a new technique for studying biomolecular interactions since the 1990s. The technical principle is to immobilize a layer of biomolecules on the surface of the sensor chip. When the sample to be measured flows over the chip surface, the molecules in the sample that interact with the biomolecules on the chip surface combine together to cause changes in the refractive index or thickness of the metal film surface. , which is finally manifested as the change of the SPR resonance angle, based on which information such as the concentration, affinity, kinetic constant and specificity of the target analyte can be obtained. SPR technology has the advantages of high sensitivity, fast detect...

Claims

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Application Information

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IPC IPC(8): G01N21/552
Inventor 苏晖崔政闫安隗玮邓绍立廉双秋苏宇梵
Owner BEIJING ZHONGLONGYICHENG TECH CO LTD
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