Method for detecting object to be detected by using surface plasmon resonance (SPR) biosensor

A biosensor and surface plasmon technology, which is applied in the field of ion resonance biosensors to detect objects to be tested, can solve problems such as easy damage to streptavidin and shedding, and achieve the effects of simplified detection and analysis of results, stable signals, and improved detection sensitivity.

Inactive Publication Date: 2012-06-27
SHENZHEN ACAD OF INSPECTION & QUARANTINE
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Problems solved by technology

However, when the streptavidin gold magnetic particles prepared by physical adsorption are washed with PBST buffer (1×PBS, containing 0.05% Tween-20), since streptavidin molecules do not contain cysteine ​​residues, , there is no strong Au-S interaction between streptavidin and gold magnetic particles, it is limited to hydrophobic interaction and electrostatic interaction, therefore, surfactant Tween-20 can easily destroy this effect and make streptavidin Almost all of the and Su have fallen off

Method used

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  • Method for detecting object to be detected by using surface plasmon resonance (SPR) biosensor
  • Method for detecting object to be detected by using surface plasmon resonance (SPR) biosensor
  • Method for detecting object to be detected by using surface plasmon resonance (SPR) biosensor

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Embodiment 1

[0053] Example 1. Using Biotin-Streptavidin-Magnetic Particles to Improve the Detection Sensitivity of SPR Biosensors

[0054] 1. Probe fixation

[0055] (1) First use 2mL 98% H 2 SO 4 Aqueous solution: 30% H 2 o 2 The bare gold film on the sensor chip J1 was rinsed with a mixed solution of aqueous solution (volume ratio 7:3), rinsed with deionized water, and dried with nitrogen gas.

[0056] (2) The thiol-modified DNA probe solution is dripped onto the surface of the sensor chip J1; the thiol-modified DNA probe sequence is as follows, obtained by biosynthesis:

[0057] 5'-HSC6-TTTTTTTTTTTTTTTT-CCATGAGAGCCTCAAGATCTGTGTTTCTTCCCT-3' (the DNA probe sequence is shown in sequence 1 in the sequence listing).

[0058] (3) Coating for 24 hours at room temperature to generate a self-assembled array of probes.

[0059] (4) Wash with deionized water and dry with nitrogen gas.

[0060] (5) Block with 6-mercaptoethanol blocking solution for 1 hour at room temperature.

[0061] 2. S...

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Abstract

The invention discloses a method for detecting an object to be detected by using an SPR biosensor. The method uses a biotin-streptavidin-magnetic particle as a signal reporting molecule which can amplify a signal of a hybridization result; therefore, the problem that marking of a probe or an object to be detected before hybridization influences natural binding of biological molecules is avoided, the requirement of improving detection sensitivity of the SPR biosensor is realized, and generated signals are stable and are easy to detect, which enables result detection and analysis to be substantially simplified. According to the invention, amplification of a detection signal is realized after hybridization, so variation and variation quantity of SPR signals before and after the detection signal is amplified can be more visually and clearly observed; meanwhile, the biotin-streptavidin-magnetic particle technology is used in the invention, and no expensive detector is needed, thereby reducing technical cost and economic cost needed for enhancing sensitivity of the biosensor.

Description

technical field [0001] The invention relates to a method for detecting an analyte with a surface plasmon resonance biosensor. Background technique [0002] SPR (Surface Plasmon Resonance) biosensor is an advanced biochemical analysis method and technology developed in recent years, and it is an analysis technology based on physical optical properties. The sensor chip technology based on SPR biosensor also emerges at the historic moment. It is a carrier of real-time signal transduction and a new technology that combines SPR sensor technology, biochip technology and molecular biology technology. At present, the SPR sensor chip mainly includes sensor chip J1, sensor chip CM5, sensor chip SA, sensor chip NTA, sensor chip HPA, and sensor chip L1. The chip structure is covered with a layer of gold film on the glass plate, and various types of chips are connected with different polymers on the surface of the gold film to form different surface environments, which is conducive to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/55
Inventor 顾大勇滕娟史蕾季明辉刘春晓赵纯中杨燕秋孙秋香赵芳欧青叶徐华徐云庆孙杰
Owner SHENZHEN ACAD OF INSPECTION & QUARANTINE
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