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Polydopamine-modification-based method for binding anti-bodies to surface of optical fiber SPR sensor

A polydopamine and sensor technology, used in instruments, scientific instruments, material analysis by optical means, etc., can solve problems such as the reduction of antibody efficiency, and achieve the effects of rapid response, simple operation, and high antibody connection efficiency.

Inactive Publication Date: 2015-11-18
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, since this method needs to treat the sensor surface with 1-ethyl-3-(3-trimethylaminopropyl)carbodiimide hydrochloride / N-hydroxysuccinimide (EDC / NHS) before antibody conjugation The carboxyl group is activated, and there is a serious hydrolysis phenomenon in this activation process, which leads to a great reduction in the efficiency of this method to link antibodies

Method used

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  • Polydopamine-modification-based method for binding anti-bodies to surface of optical fiber SPR sensor
  • Polydopamine-modification-based method for binding anti-bodies to surface of optical fiber SPR sensor

Examples

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Effect test

Embodiment 1

[0021] Clean the SPR sensor with ethanol and water, dry it with nitrogen, and set it aside. Weigh dopamine hydrochloride, dissolve it in 10 mM Tris buffer, and prepare a 2 mg / mL dopamine solution. Soak the cleaned sensor in the prepared dopamine solution, react in a shaker (120 rpm) for 30 minutes, take it out, rinse it with deionized water, blow dry with nitrogen, and the surface of the gold film is functionalized with polydopamine. Connect the polydopamine-modified sensor to the fiber-optic SPR spectrometer through SMA and other joints, and then soak it in 100 μg / mL goat anti-human immunoglobulin G in PBS, and the fiber-optic SPR spectrometer monitors the polydopamine-modified sensor in real time The resonance wavelength changes, and the active spectrum is superimposed every certain time, and the resonance wavelength of the sensor at this moment is recorded. Use the origin software to draw a graph to obtain the kinetic adsorption curve of the polydopamine-modified SPR senso...

Embodiment 2

[0025] Put the sensor with the linked antibody reaction obtained in Example 1 that has initially reached equilibrium, and put it in a 4°C incubator to continue the reaction overnight, then take it out, rinse it with PBS, dry it with nitrogen, and immerse it in a 2 μg / mL human immunoglobulin G solution , react at 37°C for 30 minutes, measure the resonance wavelength of the sensor in PBS at this time, and calculate the resonance wavelength shift value of the sensor due to the interaction of antigen and antibody; soak the sensor combined with human immunoglobulin G in 10mM NaOH for 10 minutes, and the antigen Antibody interaction is released, and the antibody on the surface of the sensor is regenerated; the regenerated polydopamine-modified sensor connected with the antibody is immersed in PBS with different concentrations of human immunoglobulin G, and the resonance wavelength of the sensor caused by each concentration of human immunoglobulin G is recorded The offset value of the...

Embodiment 3

[0028] Clean the SPR sensor with ethanol and water, dry it with nitrogen, and set it aside. Dopamine hydrochloride was weighed and dissolved in 10 mM Tris buffer to prepare a 1 mg / mL dopamine solution. Soak the cleaned sensor in the prepared dopamine solution, react in a shaker (120 rpm) for 30 minutes, take it out, rinse it with deionized water, dry it with nitrogen, and the surface of the gold film is functionalized with polydopamine. Connect the polydopamine-modified sensor to the fiber-optic SPR spectrometer through SMA and other connectors, and then soak it in PBS of 50 μg / mL goat anti-human immunoglobulin G antibody, and the fiber-optic SPR spectrometer monitors the polydopamine-modified sensor in real time. When the resonance wavelength of the sensor changes and reaches equilibrium, transfer the linked antibody reaction to a 4°C incubator to continue the reaction overnight, then take it out, wash it with PBS, dry it with nitrogen gas, and measure the resonance wavelengt...

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Abstract

The invention relates to apolydopamine-modification-based method for binding anti-bodies to surface of optical fiber SPR sensor.A rapid and simple spontaneous polymerization of dopamine on a gold membrane surface forms a gold membrane surface with polydopaminemodification.Through a Schiff base reaction carried out between quinonylsin polydopamine on the surface of the gold membrane and aminos in the antibodies, the antibody is connected to the surface of the gold membrane modified by polydopamine. According to the invention, after the antibodies are connected, the sensor resonance wavelength offset amount is two times of that of atraditional method; the sensitivity of the antigen detection is about four times of that of the antigen detection by a traditional antibody connection method, and the detection limit is about seven times lower than that a traditional antibody connection method. The sensor of the present invention can be effectively functionalized by self polymerization of dopamine, and thefunctionalizing operation is simple and the reaction is fast. Compared with the traditional antibody connection method, the antibody connection efficiency is higher, and antigen detection has a higher sensitivity. The invention has a better economic applicability and feasibility.

Description

technical field [0001] The invention relates to a method for connecting antibodies on the surface of a fiber optic SPR sensor based on polydopamine modification, which belongs to the method for surface modification and functional molecule connection of surface plasmon resonance spectrometer sensors. technical background [0002] Fiber-optic surface plasmon resonance (SPR) sensors are used to monitor molecular interactions and quantitative detection of target molecules. They have been widely used in disease diagnosis, DNA hybridization and cell behavior monitoring, environmental and food safety testing and other fields. It not only maintains the characteristics of traditional prism-type SPR sensors such as high sensitivity, no markers, and real-time detection, but also has its unique advantages, including: miniaturization, resistance to electromagnetic wave interference, and long-distance sensing. [0003] The binding efficiency of antibodies on the sensor surface plays a key...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/94G01N21/552
CPCG01N33/9413G01N21/553
Inventor 苏荣欣石瑟黄仁亮齐崴王梦凡
Owner TIANJIN UNIV
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