Colorimetric biosensor for detecting A beta<1-42> oligomer

A biosensor, oligomer technology, applied in the field of colorimetric biosensors

Active Publication Date: 2019-07-09
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is currently no method for detecting Aβ using black pho

Method used

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  • Colorimetric biosensor for detecting A beta&lt;1-42&gt; oligomer
  • Colorimetric biosensor for detecting A beta&lt;1-42&gt; oligomer
  • Colorimetric biosensor for detecting A beta&lt;1-42&gt; oligomer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Preparation of black phosphorene nano-gold composite material (BP-Au)

[0024] (1) The preparation of low-layer black phosphorus (FL-BP) adopts the ultrasonic liquid phase peeling method: accurately weigh 40mL ultrapure water and add 50mL centrifuge tube to Ar gas for 30 minutes, weigh 20mg black phosphorus into the centrifuge tube, Put the black phosphorus mixture into a cell disruptor and sonicate for 5 hours; the sonicated black phosphorus suspension is centrifuged on a high-speed refrigerated centrifuge at 2000 rpm for 10 minutes; after centrifugation, the supernatant and bottom sediment can be observed. Transfer the clear liquid to a new centrifuge tube; continue to place the centrifuge tube containing the supernatant into a high-speed refrigerated centrifuge for centrifugation, set the speed to 10000 rpm for 20 minutes; after the end of centrifugation, obvious supernatant and precipitation can be observed , Pour out the supernatant to retain the precipitate...

Embodiment 2

[0028] Example 2 Preparation of BP-Au colorimetric biosensor

[0029] (1) Prepare a 5'end sulfhydryl modified AβO aptamer TTT TTT TTT TGC CTG TGGTGT TGG GGC GGG TGC G (SEQ ID NO: 01) solution at a concentration of 10 nM.

[0030] (2) Prepare AβO solutions with different concentrations, and set the concentrations as: 0.1 nM, 1 nM, 10 nM, 100 nM, 1 μM. Take six gold-plated silicon wafers numbered 1-6 and drip 20 μL of 10 nM nucleic acid aptamer solution on the gold-plated surface respectively, and incubate for 60 min.

[0031] (3) Add 40 μL of nucleic acid aptamer solution and 80 μL of freshly prepared BP-Au to a 1.5 mL centrifuge tube, incubate for 60 minutes, and centrifuge at 8000 rpm for 5 minutes.

[0032] (4) After the incubation is completed, 10 μL of AβO solution of different concentrations are added dropwise to the gold-plated silicon wafers 1-5, and the concentration is sorted from large to small, and 10 μL of PBS solution is added dropwise to the No. 6 silicon wafer, and incu...

Embodiment 3

[0036] Example 3 Aβ 1-42 Detection of oligomers:

[0037] (1) Prepare Aβ with a concentration of 100nM separately 1-42 Monomers, oligomers, fiber bodies.

[0038] (2) After the gold-plated silicon wafer and the nucleic acid aptamer are incubated, add 10μLAβ to the silicon wafer respectively 1-42 Monomer, oligomer, cellulosic solution, incubate for 60 minutes.

[0039] (3) Add 25 μL of the prepared mixture of aptamer and BP-Au dropwise to the gold-plated silicon wafer, and incubate for 60 minutes.

[0040] (4) Clean the gold-plated silicon wafer with pure water, and perform UV-vis spectrum scanning on 4-NP.

[0041] The result is Figure 5 As shown, the reaction kinetic parameter k is obtained by first-order kinetic fitting at 400 nm. Such as Figure 5 As shown in A, when 100nM AβO is added, it has a catalytic effect on the 4-NP reaction, and the kinetic parameter of the reaction is k = 1.05x10 -5 s -1 , When the sample added is 100nM Aβ 1-42 In the case of monomer or fibrous body, th...

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Abstract

The invention discloses a colorimetric biosensor for detecting an A beta<1-42> oligomer. The colorimetric biosensor comprises a black phosphorene nano-gold composite, a gold-plated silicon wafer, andan aptamer which is specifically combined with the A beta<1-42> oligomer, wherein the 5' end of the aptamer is modified with SH; the aptamer is connected with the gold-plated silicon wafer and the black phosphorene nano-gold composite through the SH, and the aptamer is specifically combined with the A beta<1-42> oligomer; and a reaction for reducing 4-nitrophenol into 4-aminophenol through sodiumborohydride is catalyzed through the black phosphorene nano-gold composite, the catalysis effect is detected through an ultraviolet visible light spectrum, and the A beta<1-42> oligomer specifically combined with the aptamer is detected. The specificity of the A beta<1-42> oligomer can be detected under the ultraviolet visible light spectrum.

Description

Technical field [0001] This invention belongs to Aβ 1-42 The technical field of oligomer detection, specifically related to a detection of Aβ oligomers 1-42 Colorimetric biosensor. Background technique [0002] Aβ in patients with Alzheimer's disease (AD) 1-42 The concentration of oligomer (AβO) is 70 times higher than normal, so Aβ 1-42 Oligomers are the core biomarkers of AD (The Journal of Physical Chemistry C, 2017, 121(36): 20007-20015). There is currently no black phosphorene nanocomposite material to detect Aβ 1-42 Colorimetric biosensor for oligomers. Summary of the invention [0003] The purpose of the present invention is to provide a method for detecting Aβ 1-42 Colorimetric biosensor for oligomers. [0004] Another object of the present invention is to provide the application of the black phosphorene nano-gold composite material. [0005] The principle of the present invention is as follows: [0006] Such as figure 1 As shown, the black phosphorene nano-gold composite mat...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/3103G01N21/78
Inventor 孙莉萍潘伟萍
Owner XIAMEN UNIV
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