Preparation method and application of a kind of immune biosensor

A biosensor, Prussian blue technology, used in instruments, scientific instruments, measuring devices, etc., can solve the problems of time-consuming, difficult to perform on-site detection, and inconvenient to carry detection equipment, and achieve a simple and efficient preparation method and excellent sensitivity. Effect

Active Publication Date: 2019-10-11
深圳市泉晟生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods can be used for qualitative and quantitative analysis of porcine epidemic diarrhea virus, they are time-consuming, and the detection equipment is not convenient to carry, so it is difficult to perform on-site detection

Method used

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  • Preparation method and application of a kind of immune biosensor
  • Preparation method and application of a kind of immune biosensor
  • Preparation method and application of a kind of immune biosensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Preparation of immune biosensor

[0033] A method for preparing an immune biosensor, comprising the following process steps:

[0034] 1) Use a pipette gun to take 10 μL of graphene oxide dispersion solution with a concentration of 1 mg / mL and drop-coat it on the surface of the electrode substrate, dry it at room temperature, and then place it in a 0.1 mol / L potassium chloride solution, and use cyclic voltammetry to The scan rate of 100mV / s scans 20 times in the potential range of -1.5 ~ 0.5V, and the reduced graphene modified electrode is obtained;

[0035] 2) Insert the reduced graphene-modified electrode obtained in step 1) into a Prussian blue solution with a concentration of 0.1mol / L, and scan 60 laps within the potential range of 0-1.0V by cyclic voltammetry at a scan rate of 50mV / s, Form a Prussian blue deposition layer to obtain a Prussian blue / graphene modified electrode;

[0036] 3) Apply the porcine epidemic diarrhea virus monoclonal antibody d...

Embodiment 2

[0040]Embodiment 2: Determination of detection conditions of immune biosensor

[0041] (1) Effects of different pH on its detection of porcine epidemic diarrhea virus

[0042] The antibody-coated immunobiosensors were tested by cyclic voltammetry in the bottom solution of pH6.4, pH6.7, pH7.0, pH7.5 and pH8.0, and the test results were as follows: figure 1 As shown, the bottom solution with pH 7.0 is one of the optimal conditions for the immunobiosensor to detect porcine epidemic diarrhea virus with the same scanning speed and the same potential range.

[0043] (2) Effects of different base solutions on the detection of porcine epidemic diarrhea virus

[0044] The antibody-coated immunobiosensors were respectively placed in distilled water and the bottom solution constant in PBS, and tested by cyclic voltammetry. The test results are as follows: figure 2 As shown, the same scanning speed and the same potential range are obtained, and the bottom solution constant volume of PB...

Embodiment 3

[0049] Example 3: Performance testing of immune biosensors

[0050] (1) Stability

[0051] The immunosensor was stored in a refrigerator at 4°C, and retested after 30 days, and the current response was recorded. Compared with the initial signal, it had good consistency.

[0052] (2) specificity

[0053] The immunosensor is used to detect positive contrast samples containing PRRVS, CSFV, and FMDV, and the virus in the samples is detected by cyclic voltammetry, and the detection results are as follows: Figure 5 As shown, it can be seen that the current of the comparative sample drops significantly, which is obviously different from the detection result of porcine epidemic diarrhea virus.

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Abstract

The invention discloses a manufacturing method for an immunobiosensor. The manufacturing method comprises the following process steps: 1) manufacturing a reduced graphene modified electrode; 2) manufacturing a prussian blue / graphene modified electrode; and 3) dispensing a porcine epidemic diarrhea virus monoclonal antibody onto the surface of the prussian blue / graphene modified electrode, drying at the room temperature, soaking into a bovine serum albumin solution with mass concentration of 1-2%, and performing water-bath reaction for 0.5-1 hour, thereby obtaining the immunobiosensor. According to the manufacturing method disclosed by the invention, the porcine epidemic diarrhea virus monoclonal antibody is dispensed to the surface of the prussian blue / graphene modified electrode, and the bovine serum albumin solution is utilized to seal the rest of non-specific active sites on the electrode; meanwhile, important process parameters in the manufacturing method are limited, so that the manufactured immunobiosensor has excellent sensitivity, accuracy and stability. The invention further discloses application of the immunobiosensor manufactured from the manufacturing method to clinical quick detection on porcine epidemic diarrhea in non-laboratory setup.

Description

technical field [0001] The invention relates to the field of immune biosensors, in particular to a preparation method and application of an immune biosensor. Background technique [0002] Graphene is a special two-dimensional carbon nanomaterial discovered in recent years and only has a single-atom thickness. It has high electrical conductivity, large specific surface area, room temperature electron mobility, and excellent mechanical properties. The electron velocity is extremely fast at room temperature, and the electron mobility exceeds 15000cm 2 / V·S, far exceeds the transfer speed of electrons in general conductors, and its resistivity is extremely low, only 10 -8 Ω·cm, and various atoms and molecules can be adsorbed and desorbed by it, and the structure is very stable. Graphene-modified electrodes have been applied to the detection of various substances. Prussian blue, also known as iron blue, Berlin blue, etc., the molecular formula is Fe 4 [Fe(CN) 6 ] 3 , with ex...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N27/30G01N27/327G01N27/48
Inventor 李智丽赵云翔黄淑坚廖洁丹黄兴国覃丽梅刘佑明迟仕红
Owner 深圳市泉晟生物科技有限公司
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