Surface plasma resonance sensing chip and preparation method and application thereof

A surface plasmon and sensor chip technology, used in measurement devices, material analysis by optical means, instruments, etc., can solve the problems of high instrument maintenance costs, expensive CM5 chips, and long time required, and achieves short cycle times. The effect of fixed capacity and good uniformity

Active Publication Date: 2016-10-12
INST OF ELECTRONICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

This commercial CM5 chip has the advantages of good repeatability and high stability. The disadvantage is that the commercial CM5 chip is very expensive, and the price of a single chip is more than 1,000 yuan. The cost is also very high
Although the CM5 chip can be reused, once a certain antibody is immobilized on the channel, the second antibody cannot be immobilized, which lacks flexibility.
Sephadex layer preparation process (Masson J.F., Battaglia T.M., Davidson M.J., Yoon-Chang K., Prakash A.M.C., Stephen B., "Biocompatible polymers for antibody support on gold surfaces", Talanta, 2005, 67 (5 ), pp.918-925.) are also very cumbersome and take a long time, usually 5 days

Method used

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  • Surface plasma resonance sensing chip and preparation method and application thereof
  • Surface plasma resonance sensing chip and preparation method and application thereof
  • Surface plasma resonance sensing chip and preparation method and application thereof

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Experimental program
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preparation Embodiment 1

[0064] Such as figure 1 As shown, it is a schematic flow diagram of the preparation method of the present invention. Specifically, the preparation method of the surface plasmon resonance sensor chip of the present invention comprises the following steps:

[0065] (1) Cleaning the glass substrate: washing the glass substrate with concentrated sulfuric acid and deionized water successively, and drying;

[0066] (2) A chromium film with a thickness of 3-10nm is prepared on a glass substrate by sputtering or deposition, and then a gold film with a thickness of 45-55nm is prepared on the chromium film. The solid-phase carrier of the sensor chip is made into a bare gold chip;

[0067] (3) Drop the solution containing modified bovine serum albumin on the bare gold chip, so that the solution evenly covers the entire chip, continue to drop until the entire chip is soaked, seal the oscillation reaction, and use the self-assembly method to carry out the surface of the gold film. Chemi...

preparation Embodiment 2

[0072] The preparation method of the off-line immobilized small molecule detection SPR sensor chip also includes the following step (6), taking the cortisol chip as an example.

[0073] Described step (6) comprises following substep:

[0074] a) Add the mixed solution of NHS and EDC (1:1 mixing, the concentration of NHS 0.1mol / L, the concentration of EDC 0.4mol / L) on the blank chip, so that the solution evenly covers the entire chip, continue to drop until soaked The whole chip is sealed and oscillating, so that the modified bovine serum albumin immobilized on the surface of the gold film undergoes a chemical cross-linking reaction, and reacts at room temperature for 15 minutes to 1 hour, and then cleans and dries the chip;

[0075] b) Drop a cortisol conjugate solution with 0.1M acetic acid buffer as a solvent on the chip, the concentration of the conjugate is 300mg / L, so that the solution evenly covers the entire chip, continue to drop until the entire chip is soaked, seal a...

preparation Embodiment 3

[0078] The preparation method of the online immobilized small molecule detection SPR sensor chip also includes the following step (6), taking the cortisol chip as an example.

[0079] Described step (6') comprises following sub-steps:

[0080] a') Install the blank chip on the surface plasmon resonance biochemical analyzer, and fix the supporting flow cell;

[0081] b') flow into the mixed solution of NHS and EDC (1 to 1 mixing, the concentration of NHS 0.1mol / L, the concentration of EDC 0.4mol / L), so that the modified bovine serum albumin immobilized on the surface of the gold film undergoes chemical For cross-linking reaction, react at room temperature for 7 minutes to 0.5 hours, and then wash with PBS buffer;

[0082] c') flow into the cortisol conjugate solution with 0.1M acetate buffer as solvent, the concentration of the conjugate is 100mg / L, react at room temperature for 10 minutes to 1 hour, and then flow wash with PBS buffer;

[0083] d') Continuously and slowly flo...

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Abstract

The invention discloses a surface plasma resonance sensing chip and a preparation method and application thereof. The preparation method comprises the steps that firstly, a glass substrate is washed and then dried; secondly, a chromium film is prepared, a gold film is prepared on the chromium film, and a naked gold chip is obtained; thirdly, a solution containing modified bovine serum albumin is dropwise added, the whole chip is paved with the solution, the solution continues to be dropwise added until the whole chip is soaked, and after a sealed vibration reaction is carried out for 0.5-2 hours, washing and drying are carried out; fourthly, a mixed solution of NHS and EDC is dropwise added, the whole chip is paved with the mixed solution, the solution continues to be dropwise added until the whole chip is soaked, and after a sealed vibration reaction is carried out for 15 minutes to 1 hour, washing and drying are carried out; fifthly, a bromoacetic acid solution is dropwise added, the whole chip is paved with the bromoacetic acid solution, the bromoacetic acid solution continues to be added until the whole chip is soaked with the bromoacetic acid solution, after a sealed vibration reaction is carried out for 8-24 hours, washing and drying are carried out, and a blank chip is obtained. According to the preparation method, the surface plasma resonance sensing chip can be obtained within only 2 days and has better homogeneity than a traditional glucan chip which is prepared within 5 days.

Description

technical field [0001] The invention belongs to the technical field of surface plasmon sensor chip preparation and biomedical detection, and in particular relates to a surface plasmon resonance sensor chip that can be used for small molecule detection and its preparation method and application. Background technique [0002] Cortisol is the most important biochemical indicator for early detection and diagnosis of whether people suffer from depression. When people are under heavy work and life pressure, are under stress for a long time, or because of the tense pace of life, or sleep less than 8 hours a night, etc., the concentration of cortisol in their blood, urine, and saliva The level will increase or remain high for a long time. This is when the negative effects of cortisol begin to manifest as metabolic changes: elevated blood sugar, increased appetite, weight gain, depression, extreme fatigue, and more. [0003] Cortisol is a steroid hormone that regulates blood pressu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/552
CPCG01N21/553
Inventor 崔大付陈兴张璐璐徐春方李亚亭任艳飞
Owner INST OF ELECTRONICS CHINESE ACAD OF SCI
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