Function of surfactant, and signal amplification method for SPR (Surface Plasma Resonance) detection on low molecular weight substances

A surfactant and surface plasmon technology, applied in the field of bioinformatics, can solve problems such as complex experimental processes, and achieve the effect of high experimental repeatability and easy operation

Active Publication Date: 2014-04-16
苏州普芯生命科学技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conversely, if there is too little avidin, all the biotin binding sites on it will be blocked by biotinylated antibodies, so that the composite antibody cannot react with the biotin on the detection antibody, so although the avidin-biotin system is in a certain Improve the sensitivity of the detection to a certain extent, but at the same time complicate the experimental process
Moreover, these methods are rarely used in the detection and analysis of small molecules. Therefore, it is particularly necessary to find a method that is simple and easy to operate and can significantly amplify the reaction signal to improve the detection sensitivity of low molecular weight substances.

Method used

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  • Function of surfactant, and signal amplification method for SPR (Surface Plasma Resonance) detection on low molecular weight substances
  • Function of surfactant, and signal amplification method for SPR (Surface Plasma Resonance) detection on low molecular weight substances
  • Function of surfactant, and signal amplification method for SPR (Surface Plasma Resonance) detection on low molecular weight substances

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0120] Example 1 Amplification of SPRI signals by treating the substrate small molecule 4-nitrophenyl octanoate (4-Nitrophenyl octanoate) with surfactant Tween-80

[0121] 1. Immobilize 100ug / ml lipase CalB on the surface of the SPRI chip, and use 200ug / ml bovine serum albumin BSA as a negative control;

[0122] 2. Prepare PBS+0.05% Tween-80 buffer;

[0123] 3. Fix the chip on the SPRI device, and scan the intensity of the fixed positions of lipase CalB and bovine serum albumin BSA through the program selection, and pass the surface into PBS+0.05%Tween-80 buffer for baseline scanning, such as figure 1 Baseline shown;

[0124] 4. Use the prepared PBS+0.05%Tween-80 buffer to prepare the substrate small molecule 4-nitrophenyl octanoate

[0125] Diluted from 10mM of the mother solution to 0.25mM to obtain sample 1;

[0126] 5. Inject sample 1 at a binding rate of 2ul / s, such as figure 1 Combined with the curve shown;

[0127] 6. Flush with PBS+0.05% Tween-80 buffer until the ...

Embodiment 2

[0129] Example 2 Amplification of SPRI signal by treating the substrate small molecule 4-nitrophenyl octanoate (4-Nitrophenyl octanoate) with surfactant Tween-80

[0130] 1. Immobilize 100ug / ml lipase CalB on the surface of the SPRI chip, and use 200ug / ml bovine serum albumin BSA as a negative control;

[0131] 2. Prepare PBS+0.05% Tween-80 buffer;

[0132] 1. Fix the chip on the SPRI device, select the fixed positions of lipase CalB and bovine serum albumin BSA for intensity scanning, and pass the surface into PBS buffer for baseline scanning, such as image 3 Baseline shown;

[0133] 2. Dilute the substrate small molecule 4-nitrophenyl octanoate from the mother solution 10mM to 0.25mM with the prepared PBS+0.05%Tween-80 buffer solution to obtain sample 1;

[0134] 3. Pass through sample 1 at a binding rate of 2ul / s, and the binding time is 400s, such as image 3 Combined with the curve shown;

[0135] 4. Flush with PBS buffer until the baseline is stable, such as image...

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Abstract

The invention relates to the field of biological information, in particular to the function of a surfactant, and a signal amplification method for SPR (Surface Plasma Resonance) detection on low molecular weight substances. According to the method, molecules with smaller relative molecular mass are aggregated into micelles through the surfactant to form macromolecules, and the macromolecules react with enzymes, the sensitivity is very high, and the reaction speed is very fast, so that the method is suitable for quick and sensitive detection; compared with other signal amplification methods, the signal amplification method provided by the invention is on the basis of the principle that the micelles can be formed by aggregating through the surfactant, simple and convenient to operate, and high in experimental repeatability.

Description

technical field [0001] The invention relates to the field of biological information, in particular to the effect of surfactants and a method for signal amplification of low-molecular-weight substances detected by surface plasmon resonance. Background technique [0002] Surface plasmon resonance (Surface Plasmon Resonances, SPR) sensing technology is a biomolecular detection technology developed in the 1990s. The principle is: when the light is totally reflected on the surface of the prism and the metal film, an evanescent wave will be generated in the metal film, and when the evanescent wave resonates with the surface plasmon wave, the intensity of the detected reflected light will be greatly weakened. The energy is transferred from the photons to the surface plasmon, and most of the energy of the incident light is absorbed by the surface plasmon wave, so that the energy of the reflected light is drastically reduced. The corresponding incident angle at this time is the reso...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/55
Inventor 王丽红欧小敏彭开美何建安朱劲松
Owner 苏州普芯生命科学技术有限公司
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