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Disposable multi-channel electrochemical immunosensor with high sensitivity

An immune sensor and multi-channel technology, which is applied in the field of high-sensitivity disposable multi-channel electrochemical immunosensor, can solve the problems of unfavorable detection system miniaturization, high price, inconvenient operation, etc., to eliminate the interference of dissolved oxygen, low cost, The effect of easy operation

Inactive Publication Date: 2011-05-11
NANJING UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these electrode arrays are mostly prepared by photolithography and other processes, which are relatively expensive, and the usual electrochemical detection method is based on horseradish peroxidase catalyzed peroxidation reduction, which is easily interfered by dissolved oxygen in the detection solution. Nitrogen and other methods to remove oxygen are very inconvenient to operate and are not conducive to the further miniaturization of the detection system

Method used

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  • Disposable multi-channel electrochemical immunosensor with high sensitivity
  • Disposable multi-channel electrochemical immunosensor with high sensitivity
  • Disposable multi-channel electrochemical immunosensor with high sensitivity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: in combination with figure 1 , Preparation of glucose oxidase functionalized nanoprobes

[0032] Multi-walled carbon nanotubes in 3:1H 2 SO 4 / HNO 3 Ultrasonic treatment in medium for four hours, washed with water until neutral, and dried to obtain carboxylated carbon nanotubes. Next, disperse 0.75mg of pretreated carbon nanotubes in 1.5mL of 0.20% polyelectrolyte PDDA containing 0.5M NaCl, sonicate for 30 minutes, discard excess PDDA by high-speed centrifugation, and wash three times with water to obtain PDDA functionalized carbon nanotubes. The PDDA functionalized carbon nanotubes were dispersed in 9.0 mL of 13-nm gold colloidal nanoparticles, stirred for 20 minutes, centrifuged, and washed with water three times to obtain light purple carbon nanotube composites loaded with gold nanoparticles. Next, disperse the complex in 2.5mL 50mM pH 9.0 Tris-HCl solution, add 1.9mL 2mg / mL glucose oxidase and 75μL 0.5mg / mL secondary antibody, stir the reaction for...

Embodiment 2

[0033] Embodiment 2: in combination with figure 2 , Fabrication of Disposable Multichannel Immunosensors

[0034] At room temperature, under constant stirring, add 6.25mM FeCl to 16mL 2 , 0.4% polyelectrolyte PDDA and 0.15% chitosan solution, slowly drop 4mL 25mM K 3 Fe[(CN) 6] solution, the solution gradually turns dark blue, thereby generating the desired Prussian blue complex. The screen-printed electrode array was pre-oxidized with 0.1mol / L dilute sulfuric acid at 1.3V constant potential for 120s, washed and dried, and then 1 μL of Prussian blue complex was added dropwise on the working electrode, left to dry at room temperature and then adsorbed at room temperature for 13 -nm gold nanoparticles for 6 hours, washed successively with rinse solution and pH 7.0 phosphate buffer solution, and dried in the air. On the surface of different Prussian blue complexes / gold nanoparticles modified working electrodes, add 0.5 μL of 0.5 mg / mL different capture antibodies dropwise, a...

Embodiment 3

[0035] Embodiment 3: in combination with image 3 , a multichannel immunoassay method

[0036] (1) On the prepared multi-channel immunosensor, drop 10 μL of different concentrations of standard antigen or serum to be tested, incubate at room temperature for 40 min, and wash with washing solution;

[0037] (2) Add 10 μL of 5-fold diluted nanocomposite probe dropwise and incubate for 40 minutes, then wash with washing solution;

[0038] (3) A detection solution containing 10 mM glucose was added dropwise, and differential pulse voltammetry was performed in the potential range from 0.30 V to -0.20 V, with a pulse amplitude of 50 mV and a pulse width of 50 ms. According to the recorded electrochemical signals, the working curves of different analyte components are obtained, and multi-channel simultaneous determination of the analyte components is performed.

[0039] (4) The multi-channel immunosensor is for one-time use.

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Abstract

The invention relates to a disposable multi-channel electrochemical immunosensor with high sensitivity. A prussian blue composite, nanogold and a capture antibody are assembled layer by layer on a disposable printing electrode array to obtain a multi-channel immunosensor. Enzyme and secondary antibody in great proportion are assembled on a carbon nano tube loading gold nanoparticles and a novel glucose oxidase functional nano composite probe is designed for sandwich immunoassay. The nano composite probe is combined with the multi-channel immunosensor to realize double signal amplification and high sensitive immunodetection of protein. The prussian blue, as electronic transmission media, catalyzes and reduces hydrogen peroxide generated by glucose oxidation under the catalysis of glucose oxidase on oxygen so as to obtain current signal. The method avoids the interference of dissolved oxygen in the detection solution and deoxidization is not required in process of amperometric detection. The invention has the advantages of wide detection concentration range, good repeatability, accurate results and the like and has a certain clinical application value.

Description

1. Technical field [0001] The invention is a high-sensitivity disposable multi-channel electrochemical immunosensor. It uses the signal amplification and specific recognition of the glucose oxidase functionalized nanocomposite probe, combined with the disposable multi-channel electrochemical immunosensor prepared by the screen-printed electrode array, to perform high-sensitivity electrochemical immunoassay. 2. Background technology [0002] As a highly selective and sensitive analytical method, immunoassay has been widely used in clinical diagnosis, environmental monitoring, food safety and other fields. Immunoassays are divided into homogeneous and heterogeneous immunoassays, the latter being widely used because of higher sensitivity. Compared with immunoassay methods based on radiation analysis, fluorescence, chemiluminescence, electrochemiluminescence, surface plasmon resonance and other analytical techniques and traditional enzyme-linked immunoassay methods, electrochem...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/535
Inventor 鞠熀先严枫赖国松钟丹秋
Owner NANJING UNIV
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