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ECL sensing chip for rapidly detecting cysteine proteinase-3 as well as preparation method and application of ECL sensing chip

A sensor chip and luminescent material technology, applied in the field of bioelectrochemical sensors, can solve the problems of lack of performance electrochemiluminescent reagents, etc., achieve high sensitivity and stability, and broad application prospects

Active Publication Date: 2021-10-22
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the lack of electrochemiluminescent reagents with good performance and rationally designed biosensing mechanisms, there are few reports on the detection of Caspase-3 activity using this method.

Method used

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  • ECL sensing chip for rapidly detecting cysteine proteinase-3 as well as preparation method and application of ECL sensing chip
  • ECL sensing chip for rapidly detecting cysteine proteinase-3 as well as preparation method and application of ECL sensing chip
  • ECL sensing chip for rapidly detecting cysteine proteinase-3 as well as preparation method and application of ECL sensing chip

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preparation example Construction

[0034] The invention provides a preparation method of luminescent material Ru-PEI@PCN-333(Al), which comprises the following steps: [Ru(bpy) 2 (mcpbpy)] 2+ Mix with polyethyleneimine for covalent bonding reaction to get Ru-PEI complex;

[0035] The Ru-PEI complex and PCN-333(Al) were mixed and stirred for 24 hours to obtain the luminescent material Ru-PEI@PCN-333(Al).

[0036] The preparation method of the Ru-PEI complex of the present invention preferably comprises: [Ru(bpy) 2 (mcpbpy)] 2+ Mix with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) for 20 to 40 minutes, and then mix the obtained mixture with Polyethyleneimine (PEI) was continuously mixed for 2-4 hours to obtain the Ru-PEI composite. In the present invention, the [Ru(bpy) 2 (mcpbpy)] 2+ Preferably by [Ru(bpy) 2 (mcpbpy)] Cl 2 Provided that the [Ru(bpy) 2 (mcpbpy)] Cl 2It is preferred to mix and stir with EDC and NHS for 30 minutes, so that the carboxyl gr...

Embodiment 1

[0064] 1. Synthesis of PCN-333(Al)

[0065] 50mg H 3 TATB and 200 mgAlCl 3 ·6H 2 O was dissolved in 10 mL of DMF, then 1.0 mL of trifluoroacetic acid was added for ultrasonic dissolution, heated in an oven at 135 °C for 48 h, cooled to room temperature and then centrifuged to obtain a white precipitate, which was washed three times with DMF to obtain a white powder sample PCN-333 (Al ).

[0066] 2. Synthesis of Ru-PEI@PCN-333(Al)

[0067] 14.8mg [Ru(bpy) 2 (mcpbpy)] Cl 2 Dissolve in 4mL ultrapure water, add 57.6mg EDC and 11.6mg NHS under vigorous stirring, and stir for 30min to activate the carboxyl group. Then 2 mL of 1% PEI (Mw=1800) solution was added and stirred for 2 h to obtain a Ru-PEI complex. Then 10mg of negatively charged PCN-333(Al) was added and stirring was continued for 24h. The obtained mixed solution was dialyzed with a dialysis bag (8000-14000Da) for 24 hours and concentrated by centrifugation. The final product Ru-PEI@PCN-333(Al) was dispersed in 1 ...

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Abstract

The invention provides an ECL sensing chip for rapidly detecting cysteine proteinase-3 as well as a preparation method and application of the ECL sensing chip, and relates to the technical field of bioelectrochemical sensors. The preparation method comprises the following steps: taking PCN-333(Al) as a carrier of a luminous reagent Ru, and covalently binding carboxyl Ru and a co-reactant PEI to prepare a Ru-PEI@PCN-333(Al) composite material; modifying the composite material and AuNPs on the surface of an SPE working electrode by using CS as an adhesive; and modifying the ferrocene polypeptide on the electrode through an Au-S bond to construct the ECL sensing chip. The ECL signal of the Ru is quenched due to approaching of the Fc; and when the Caspase-3 is added, the ECL signal is recovered. By utilizing the sensing chip, the activity of Caspase-3 in the cell apoptosis process can be effectively monitored, and the sensing chip has a wide application prospect in the aspects of drug curative effect evaluation, cancer treatment monitoring and the like.

Description

technical field [0001] The invention belongs to the technical field of bioelectrochemical sensors, and in particular relates to an ECL sensor chip for rapid detection of cysteine ​​protease-3, a preparation method and application thereof. Background technique [0002] Apoptosis is a gene-mediated cell suicide behavior, which plays an important role in maintaining the homeostasis of the organism's environment and removing abnormal cells. Abnormal apoptosis process can lead to many serious diseases such as cancer and neurodegenerative diseases. Caspases are a class of aspartic acid-specific cysteine ​​proteases closely related to apoptosis. Caspase-3 is the most important and most studied protein in the Caspase family, and it is at the core of the apoptosis network. The level of apoptosis of cells can be reflected by the expression of Caspase-3. Therefore, monitoring the activity of Caspase-3 is of great significance for understanding its role in the process of apoptosis, a...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/48G01N21/76
CPCG01N27/3277G01N27/3278G01N27/48G01N21/76
Inventor 宋大千马品一孙颖曹彦波罗维巍
Owner JILIN UNIV
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