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
View PDF8 Cites 3 Cited by
  • 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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 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...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Concentrationaaaaaaaaaa
Login to view more

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/3103G01N21/78
Inventor 孙莉萍潘伟萍
Owner XIAMEN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products