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Biosensor for detecting PSA, preparation method and application thereof

A biosensor, capture probe technology, applied in instruments, measuring devices, scientific instruments, etc., can solve the problems of low sensitivity and poor specificity, and achieve the effects of high sensitivity, low cost and improved sensitivity

Active Publication Date: 2021-05-28
SHANGHAI UNIV OF ENG SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to overcome the defects of low sensitivity and poor specificity of existing PSA electrochemical immunosensors, and provide a PSA electrochemical sensor with high sensitivity and good specificity

Method used

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  • Biosensor for detecting PSA, preparation method and application thereof
  • Biosensor for detecting PSA, preparation method and application thereof
  • Biosensor for detecting PSA, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] A method for preparing a biosensor for detecting PSA, the steps are as follows, and the flow chart is as follows figure 1 Shown:

[0073] (1) Clean the glassy carbon electrode:

[0074] The glassy carbon electrode (Φ=2mm) is sequentially coated with 0.3μm and 0.05μm Al 2 o 3 Polish the surface of the electrode into a mirror surface with polishing powder, then ultrasonically clean it with absolute ethanol and deionized water for 5 minutes, and dry it with nitrogen gas for later use;

[0075] (2) Add 12 μL of MoS at a concentration of 20 μg / mL 2 The suspension was added dropwise to the surface of the cleaned glassy carbon electrode, and incubated at 37°C for 40min;

[0076] (3) Add 12 μL of the mixed solution of capture probe ssDNA and polyC dropwise to the surface of the modified electrode, and incubate at room temperature for 10 h. The concentrations of capture probe ssDNA and polyC in the mixed solution of capture probe ssDNA and polyC are respectively 1μM and 1μM...

Embodiment 2～5

[0081] A method for preparing a biosensor for detecting PSA is basically the same as in Example 1, except that in steps (2) to (4), the dispersion (i.e. corresponding to MoS 2 Suspension, mixed solution of capture probe ssDNA and polyC, and SiO containing modified ssDNA labeled with methylene blue 2 Particle suspension), the addition amount, incubation temperature and time, the difference is shown in the following table, in the table, A is MoS 2 The amount of suspension added, the unit is μL, B and C are the temperature and time of incubation in step (2), the unit is ℃, min respectively, D is the added amount of the mixed solution of capture probe ssDNA and polyC, the unit is μL, E is the incubation temperature in step (3), in °C, F is SiO containing modified ssDNA and labeled with methylene blue 2 The amount of particle suspension added, the unit is μL, G is the temperature incubated in step (4), the unit is °C;

[0082] A B C D E F G Example 2 10 40...

Embodiment 6

[0084] For the application of the biosensor that detects PSA, its steps are as follows:

[0085] Establish a standard curve for PSA detection:

[0086] Take out the biosensor for detecting PSA prepared in Example 1, add 12 μL of PSA with different concentrations dropwise successively, incubate at room temperature for 2 h, then wash with PBS solution, blow dry with nitrogen, and use square wave voltammetry in PBS solution Investigate the change of current, and use it as the detection result, which can be used to make a standard curve for PSA detection. In SWV scanning, the pH of the scanned PBS solution is 7.4, the concentration is 10mM, and the scanning voltage range is -0.6~0V.

[0087] The sensor detects the change of the CV diagram signal of the target PSA as follows: figure 2 As shown, the SWV signal diagram obtained by detecting PSA with different concentration gradients is shown in Figure 7As shown, the PSA concentrations from a to k are 0fg / mL, 1fg / mL, 10fg / mL, 100f...

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Abstract

The invention discloses a biosensor for detecting PSA, a preparation method and application thereof. The biosensor comprises a working electrode, the working electrode comprises a glassy carbon electrode, the surface of the glassy carbon electrode is coated with a MoS2 layer, a signal marking segment and a capture probe ssDNA specifically bound with PSA; the capture probe ssDNA is fixed on the MoS2 layer, the signal marking segment is SiO2 particles containing modified ssDNA and marked with methylene blue, and the signal marking segment is connected with the capture probe ssDNA through the modified ssDNA. The biosensor disclosed by the invention is high in sensitivity, low in PSA detection limit, convenient to detect, good in specificity and good in application prospect; the method has the advantages of simple process, mild conditions, low cost and great application prospects.

Description

technical field [0001] The invention belongs to the technical field of novel functional materials and biosensing detection, relates to a biosensor for detecting PSA and its preparation method and application, in particular to a biosensor for supersensitive detection of PSA based on nano-signal amplification and its preparation methods and applications. Background technique [0002] PSA (prostate specific antigen, prostate specific antigen) is a glycoprotein directly secreted by prostate epithelial cells into the prostate duct system. The PSA content in the serum of normal people is generally not higher than 4ng / mL, but when the prostate cancer occurs, the PSA content in the serum will rise sharply. Therefore, PSA has become a biomarker of prostate cancer, and it is also considered to be the most organ-specific marker among tumor markers. In recent years, the initial screening of PSA as the only marker of prostate cancer has been internationally recognized and has been clin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/30G01N27/38
CPCG01N27/3278G01N27/308G01N27/3277G01N27/38Y02A50/30
Inventor 严若鸿鲁娜张敏李洁陆张璐史雪荣
Owner SHANGHAI UNIV OF ENG SCI
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