Construction of early tumor high-throughput electrochemical luminescence detection method

A luminescence detection and electrochemical technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc. problem, to achieve the effect of strong designability, low price and low toxicity

Active Publication Date: 2015-03-25
LINYI UNIVERSITY +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The onset of the tumor is hidden, the symptoms are not obvious, the symptoms and imaging examination are n

Method used

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  • Construction of early tumor high-throughput electrochemical luminescence detection method
  • Construction of early tumor high-throughput electrochemical luminescence detection method
  • Construction of early tumor high-throughput electrochemical luminescence detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example One Combination figure 1 , figure 2 and image 3 , Preparation of iridium complex-filled mesoporous silicon / nano-gold composite signaling probes

[0022] 1.0mL concentration is 2.5 × 10 ?4 M [(ppy) 2 Ir(dcbpy)] + PF 6 ? ( figure 1 ) was added to the same volume of 1.0 mg / mL mesoporous silicon solution and stirred overnight at 37°C, then 1.5 mL of 3-mercaptopropyltrimethoxysilane was added and stirred for 2 hours, then the nano-gold solution was added. Nanogold is assembled onto the surface of mesoporous silicon through the combination of gold-mercapto covalent bonds to form a stable iridium complex-filled mesoporous silicon / nanogold complex signal probe. Take 1mL of the above nanocomposite and add 1mL of 0.02mg / mL antibody (secondary antibody, 0.01M PBS, pH=7.4) was incubated at room temperature for 2 h. Centrifuge, wash with 0.0 1M PBS with pH=7.4, then dissolve the resultant in 1 mL of 1% BSA, incubate for 1 h, and wash by centrifugation. The fina...

Embodiment 2

[0024] Example two combination Figure 4 , the development of flexible array microchip electrodes

[0025] Mix polydimethylsiloxane (PDMS) with an appropriate amount of curing agent at a mass ratio of 9:1, and form a transparent electrode base material after curing. Make a hollow plexiglass plate (the hollow part is in the shape of an electrode), and cover it tightly on the PDMS sheet. will contain 1% HAuCl 4 , 200 g L -1 KHCO 3 and 2% glucose (volume ratio: 2:1:1) electroless gold-plating solution was injected into the above hollowed out part, after the air bubbles were exhausted, it was left to stand for 3 hours under dark conditions, and then the gold-plating solution was removed to form a layer of texture on the surface of PDMS Uniform gold nanofilms. The nano-gold layer has excellent electrical conductivity. At the same time, with the flexibility of the substrate PDMS material, the obtained flexible microchip electrode has unique advantages in preparation, storage, ...

Embodiment 3

[0027] Example three combination Figure 5 , High-sensitivity, high-throughput electrochemiluminescent tumor marker detection based on novel signaling probes

[0028] On the surface of the designed flexible microchip, the bionic sensing interface is constructed by using the interface modification technology. Add 10 μL of primary antibody (100 μg L -1, 0.01 M pH 7.4 PBS) onto the surface of the electrode and incubated at 4 °C for 12 h, and the unbound antibody was removed by washing with buffer. Then the prepared electrode was immersed in 1 % (w / w) BSA solution for 1 h at room temperature to seal the possible active sites and non-specific adsorption on the electrode surface, and finally the electrode was washed several times with buffer solution. The modified electrodes were stored in a refrigerator at 4 °C until use. Soak the prepared immune electrode in 100 μL of tumor marker solutions of different concentrations, incubate at 37 °C for 40 min, take it out and wash it sever...

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Abstract

The invention constructs a new method of high-sensitivity high-throughput electrochemical luminescence tumor marker detection based on the development of iridium complex molecule flexible array microchip electrode having high electrochemical luminescence efficiency. According to the method, massive iridium complex molecules with high electrochemical luminescence efficiency are loaded into meso-porous silicon which serves as an electrochemical luminescence signal probe; gold nano-particles are used as a carrier of a tumor marker antibody to improve electron transfer of a medium, so that electrochemical luminescence signals can be improved; and by developing the high-throughput microchip electrode and constructing a sensing interface on the surface of a multi-array microchip electrode, on the basis of specific identification between antigen and antibody and in combination of the signal amplification technology of nano-particles and excellent electron transfer property high-sensitivity, high throughput and accurate detection on the tumor marker can be realized by means of the excellent electrochemical luminescence property of a iridium complex filled meso-porous silicon/nano-gold compound signal probe of which the surface is modified by secondary antibody. The method can be used for comprehensively screening early tumor diseases of mass people by means of the advantages of simpleness, quickness, low price, no invasion and the like.

Description

technical field [0001] The invention relates to a preparation method of an electrochemiluminescent nanometer probe and a flexible array microchip electrode capable of simultaneously detecting multiple tumor markers or multiple samples. Background technique [0002] Tumor is one of the major diseases that threaten human health and life in the world. The prevention and treatment policy of tumor diseases in our country is "prevention first, combined with prevention and treatment, focusing on three early stages". Early screening and diagnosis are the key to tumor treatment. At present, the screening of tumor diseases in clinical medicine mainly relies on clinical manifestations, imaging examinations, serological examinations, biopsy and other means. The onset of the tumor is hidden, the symptoms are not obvious, the symptoms and imaging examination are not sensitive to the diagnosis of early tumors, and it is still impossible to diagnose early tumors in time. At present, the c...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/574
Inventor 周宏刘静张宁波张书圣
Owner LINYI UNIVERSITY
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