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An electrochemical immunosensor and its preparation and application

An immunosensor and electrochemical technology, applied in the field of electrochemistry, can solve the problems of immature research and application, relatively few literatures, etc.

Inactive Publication Date: 2016-01-13
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In recent years, this technology has been gradually applied to the detection of mycotoxins in food, but there are relatively few literatures, especially the research and application of electrochemical biosensor detection technology for OTA in food is still immature

Method used

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  • An electrochemical immunosensor and its preparation and application
  • An electrochemical immunosensor and its preparation and application
  • An electrochemical immunosensor and its preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Example 1 Preparation of electrochemical immunosensor

[0084] 1. Materials and methods

[0085] 1.1 Materials

[0086] Ochratoxin A-bovine serum albumin conjugate (OTA-BSA), OTA monoclonal antibody (anti-OTA) were purchased from Beijing Huaan Maike Biotechnology Co., Ltd., alkaline phosphatase-labeled horse anti-mouse IgG (H +L) purchased from Vector Laboratories, USA, OTA standard, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), N-hydroxysuccinimide (NHS), α -Naphthyl phosphate (α-NP) and chitosan (CS) were purchased from Sigma-Aldrich Company in the United States, carboxylated single-walled carbon nanotubes were purchased from Shenzhen Nanoport Co., Ltd., methanol, acetone and other reagents were purchased from Chongqing Maoye Chemical Reagent Co., Ltd.

[0087] 1.2 Testing instruments

[0088] The CHI660D electrochemical workstation is a product of Shanghai Chenhua Instrument Company.

[0089] 1.3 Detection principle

[0090]A layer of carboxylated single...

Embodiment 2

[0106] Embodiment 2 Characterization and inspection of electrochemical immunosensor

[0107] 1. Characterization of SWNTs-COOH / CS composite film

[0108] The SWNTs-COOH / CS composite material suspension obtained in Example 1 is scanned and observed with a S-300N scanning electron microscope, as shown in figure 1 a and 1b show that the carbon tubes in the nanocomposite are in obvious tubular shape and dispersed uniformly in the solution.

[0109] Drop-coat SWNTs-COOH / CS on the surface of the glassy carbon electrode, and after drying at room temperature to form a film, a uniform layer of film-like substance can be seen on the surface of the electrode: SWNTs-COOH / CS / GCE.

[0110] 2. Electrochemical characterization of different modified electrodes

[0111] Such as figure 2 Shown is the square wave voltammetry response curve of different modified electrodes in 5mM potassium ferricyanide solution in each step:

[0112] a is bare GCE;

[0113] b is SWNTs-COOH / CS / GCE;

[0114] ...

Embodiment 3

[0127] Example 3 Optimization of Electrochemical Immunosensor and Its Conditions of Use

[0128] We also made several important conditions during the experiment, namely the final concentration of SWNTs-COOH in the SWNTs-COOH / CS composite suspension, the concentration of OTA-BSA, the concentration of anti-OTA, the dilution ratio of AP-anti-antibody, The several determination conditions of α-NP concentration were further optimized. For each condition, five points were selected from low concentration to high concentration to carry out a series of experiments.

[0129]1. In order to investigate the influence of the final concentration of SWNTs-COOH in the SWNTs-COOH / CS composite suspension on the electrochemical immunosensor, different concentrations (0.5, 1.0, 2.0, 4.0, 6.0mg / ml ) SWNTs-COOH / CS composite suspension to construct an electrochemical immunosensor. The results show that the final concentration of SWNTs-COOH is too low, its film-forming properties are poor, and it ca...

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Abstract

The invention provides an electrochemical immunosensor for detecting ochratoxin A. The electrochemical immunosensor comprises a work electrode, a reference electrode and a counter electrode, wherein the work electrode is prepared by that the surface of a base electrode is first modified by a carboxylated single-walled carbon nanotube / chitosan composite and then fixed with an ochratoxin A and albumin bovine serum conjugate. When the ochratoxin A is detected by the sensor, the concentration of the ochratoxin A is in a linear relationship in a range of 0.01-100 ng / ml, the linear equation is Y = 6.3155E-6-2.89458E-6X, the relevance coefficient is 0.99984, and the minimum detection limit is 0.004 ng / ml. Compared with the prior art, the sensor has the advantages of high sensitivity, great specificity, low cost, simplicity and convenience for operation and short detection period, is suitable for the measurement of actual samples and is expected to have actual application values.

Description

technical field [0001] The invention relates to the field of electrochemistry, in particular to an electrochemical immunosensor and its preparation and application. Background technique [0002] Ochratoxin is a secondary metabolite produced by several bacteria of the genus Aspergillus and Penicillium, which has teratogenic, carcinogenic, mutagenic, immunological toxicity, etc. The International Agency for Research on Cancer (IARC) classifies ochratoxin as 2B Carcinogens are divided into compounds with similar structures such as ochratoxin A, B, and C, among which ochratoxin A (OTA) is the most toxic. OTA-producing bacteria are widely distributed in nature, and various crops and foods such as grains, grapes and wine, Chinese herbal medicines, soybean products, beer, and tea can be contaminated by OTA. In recent years, the harm of ochratoxin has attracted great attention from all over the world, and many countries have established the highest allowable amount for it. [0003...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N27/26
CPCG01N33/561G01N33/56911
Inventor 邱景富李朝睿张弦
Owner CHONGQING MEDICAL UNIVERSITY
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