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Photoelectrochemical immunosensor for detecting S100B and preparation method thereof, and application of photoelectrochemical immunosensor

An immunosensor and photoelectrochemical technology, applied in the field of biomedical detection, to achieve good stability and sensitivity, easy operation and rapid response

Pending Publication Date: 2020-01-17
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, while maintaining C 60 The unique optoelectronic properties and endowing them with friendly biological interfaces are still very problematic and challenging

Method used

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  • Photoelectrochemical immunosensor for detecting S100B and preparation method thereof, and application of photoelectrochemical immunosensor
  • Photoelectrochemical immunosensor for detecting S100B and preparation method thereof, and application of photoelectrochemical immunosensor
  • Photoelectrochemical immunosensor for detecting S100B and preparation method thereof, and application of photoelectrochemical immunosensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Photoactive material C 60 Preparation of @PCN-224

[0041] (1) Synthesis of PCN-224: Accurately weigh 30mg ZrOCl with an analytical balance 4 ,20mg H 2 TCPP, 1080mg of benzoic acid was dissolved in 3.2mL of DMF, and the above solution was heated and reacted in a round bottom flask at 120°C for 6h, and finally washed with DMF and ethanol for several times, and dried.

[0042] (2)C 60 Synthesis of @PCN-224 complex: Weigh 1mg PCN-224, prepare 1mg / mL dispersion, solvent is ethanol; weigh 1mg C 60 , prepare a 1mg / mL dispersion, and the solvent is DMF. Measure PCN-224 dispersion liquid, C 60 Dispersion solution 1mL each, mixed ultrasonic 1h, stirred for 12h, can be prepared C 60 @PCN-224 Complex, C 60 @PCN-224 was dispersed in ethanol at a concentration of 1mg / mL and stored for later use.

Embodiment 2

[0044] Preparation of Nb82@CNQDs

[0045] (1) CNQDs were synthesized according to Hot-Tailoring of Carbon Nitride Dots with Red-Shifted Photoluminescence for Visual Double Text Encrypt and Bioimaging (Chem. Eur. J. 2019, 25, 10188-10196).

[0046] (2) Add 10 μL of 50% glutaraldehyde aqueous solution to 900 μL CNQDs aqueous solution (1 mg / mL), and sonicate for 2 hours in the dark. Then, 100 μL of Nb82 (100 μg / mL) in PBS (0.01 M, pH=7.4) was added and incubated at 4° C. for 12 h, so that Nb82 and CNQDs fully reacted. Finally, 10 μL of 1% BSA 0.01M PBS (pH=7.4) solution was added to the above solution, blocked for 30 min at 37 °C, and the prepared Nb82@CNQDs were stored in a 4 °C refrigerator for later use.

Embodiment 3

[0048] Preparation method of photoelectric immunosensor

[0049] according to figure 1 The steps shown assemble the sensor:

[0050] (1) The signal layer is fixed

[0051] To 0.25cm 2 Add 10 μL 1mg / mL C dropwise on the ITO 60 @ PCN-224 dispersion (embodiment 1), stay overnight at room temperature, wash with deionized water after drying, and dry to obtain C 60 @PCN-224 / ITO substrate electrode.

[0052] (2) Anchor recognition molecules

[0053] Add dropwise 10 μL of the mixed solution of EDC (100 mM) and NHS (50 mM) to the C obtained in step (1). 60 @PCN-224 / ITO substrate electrode, placed at room temperature for 1-2h to activate C 60 For the carboxyl groups on the surface of @PCN-224, wash with PBS, add 10 μL of 10 μg / mL Nb9 dropwise to the above electrode, and incubate at 4 °C for 12 hours to make it fully anchored and loaded on the surface of the electrode material, and then wash it with 0.01M PBS (pH=7.4) The solution washes away unbound Nb9.

[0054] (3) Non-specif...

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Abstract

The invention discloses a photoelectrochemical immunosensor for detecting S100B and a preparation method thereof, and application of a photoelectrochemical immunosensor. The photoelectrochemical immunosensor is prepared by covalently binding Nb9 to a substrate electrode modified with a C60@PCN-224, the S1OOB is specifically bound the Nb9 through an immunoreaction, and Nb82@CNQDs is specifically bound onto the S100B. The double-antibody sandwich type photoelectrochemical immunobiosensor is constructed by utilizing the principle that a quenching agent CNQDs has a very good quenching effect on C60@PCN-224 on an electrode substrate, and is used for detecting S100B. Compared with traditional optical methods and the like, the photoelectrochemical immunosensor has the characteristics of simplicity and convenience in operation, low technical requirements, quick response, low cost, easiness in microminiaturization and the like, and plays an important role in medical diagnosis.

Description

technical field [0001] The invention belongs to the field of biomedical detection, and in particular relates to a photoelectrochemical immunosensor for detecting S100B, a preparation method and application thereof. Background technique [0002] S100B is an acidic calcium-binding protein that is mainly expressed and secreted by astrocytes. The level of serum S100B in healthy people is low, and the level of serum S100B in patients with brain injury is high. Therefore, S100B is considered as an emerging biomarker of brain injury and BBB function. At present, the detection of S100B generally adopts enzyme-linked immunosorbent assay (ELISA). Although ELISA kits have the advantages of wide application and rapid development, they are expensive and time-consuming in actual detection. Therefore, it is of great significance to develop lower-cost, more practical, and more efficient detection strategies for S100B. However, so far, there are limited innovative methods for S100B detec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N27/327
CPCG01N33/6893G01N27/3275
Inventor 沈艳飞周晴张袁健
Owner SOUTHEAST UNIV
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