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A gold/quantum dot nanoprobe for the detection of active ricin in complex matrices and its application

A technology of ricin, nanoprobes, applied in the direction of nanotechnology for sensing, nanotechnology for materials and surface science, measuring devices, etc.

Active Publication Date: 2019-12-03
BEIJING CENT FOR DISEASE PREVENTION & CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problems in the above-mentioned prior art, the present invention researches and improves how to quickly detect active ricin in complex matrices, and eliminates the false positive problem that often occurs in general methods; further, in practical applications, For emergencies, ricin can be easily detected on-site without the need for large and complex equipment, ideally providing a visual and rapid detection method for ricin

Method used

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  • A gold/quantum dot nanoprobe for the detection of active ricin in complex matrices and its application
  • A gold/quantum dot nanoprobe for the detection of active ricin in complex matrices and its application
  • A gold/quantum dot nanoprobe for the detection of active ricin in complex matrices and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Preparation and characterization of ricin-capturing agent magnetic microspheres (MB@P(C-H)-mAbricin)

[0075] Synthesis of 3-((3-Aminopropylmethacrylate)dimethylammonium)-propionate (CBMAA)

[0076] The synthetic method can refer to the literature (Banerjee, I., Pangule, R.C., Kane, R.S., Adv Mater2011, 23, 690-718), and the specific steps are as follows: in a round bottom flask, 19.4g DMAPMA was dissolved in 100mL of anhydrous THF, and stirred At 0°C, anhydrous THF solution containing 11.5 g propiolactone was added dropwise, reacted for 1 h under an argon atmosphere, and then placed in a refrigerator at 4° C. for 24 h. The resulting white precipitate was filtered, washed with anhydrous THF, and dried in vacuo to obtain 17g CBMAA with a yield of 60%. The product structure was obtained by 1 HNMR identification. The synthetic formula is as follows:

[0077]

[0078] Synthesis of Propyl-4-(trimethoxysilyl)benzyl-carbonotrithioate (CTA)

[0079] RAFT ...

Embodiment 2

[0087] Example 2 Capability of ricin capture agent magnetic microspheres (MB@P(C-H)-mAbricin)) to ricin

[0088] A series of standard ricin from 10-50μg / mL was added to 1.0mL, 50mg / mL ricin capture agent magnetic microsphere solution. After 2 hours of incubation, the equilibrium concentration of ricin in the supernatant was determined by the BCA protein concentration assay. In addition, the binding kinetics between ricin and the ricin magnetic absorbent was measured by adding 50 mg of MB@P(C-H )-mAbricin was mixed with 50μg ricin in 1.0mL PBS buffer solution for determination. At incubation times ranging from 20 to 120 minutes, ricin concentrations in the supernatants were determined by the BCA protein method. The equilibrium adsorption capacity (Q) was calculated by the following equation:

[0089] Q=(C 0 -C e )·V·m -1 10 3 (mg / g)

[0090] C 0 (μg / mL) represents initial ricin concentration in PBS, C e (μg / mL) indicates the equilibrium concentration of ricin in the su...

Embodiment 3

[0095] Example 3 Assembly and characterization of gold / quantum dot nanoprobes (AuNPs / QDs)

[0096] The single-strand deoxyribonucleotides (ssODN) used in this example are P1(a), P2(a), and P3(a) in Table 1, respectively.

[0097] Spherical gold nanoparticles (AuNPs) were obtained by seed growth method, the method can refer to literature (Bastús, N.G., Comenge, J., Puntes, V., Langmuir 2011, 27, 11098–11105), with a diameter of about 60nm. The concentration of the AuNPs gel after 6 growths was 1.9×10 11 (NP / mL). In order to stabilize the gold nanoparticles, the BSPP of 15mg was added to the 100mL gold nanoparticle sol under stirring, and the reaction was at least 10 hours. The resulting BSPP-AuNP sol was centrifuged, the supernatant was removed, and then dispersed into 10 mL of ultrapure water. P1-modified gold nanoparticles (P1-AuNPs) were obtained through the strong binding between gold particles and thiols. The specific steps were to mix 50 μL of 0.1 mmol / L P1(a) with 5...

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Abstract

The invention discloses a gold / quantum dot nanoprobe for detecting active ricin in a complex matrix and an application thereof. The gold / quantum dot nanoprobe is to utilize deoxyribonucleotide single-strand (ssODN) modified gold nanoparticles and quantum dots to form deoxyribonucleotide double strands through base pairing hybridization, and gold nanoparticles and Quantum dots assembled as core-satellite nanoprobes. The invention uses the gold / quantum dot nano probe to detect active ricin, the detection limit is 7.46ng / mL, the accuracy is high, the reliability is good, and no large-scale equipment and complex operation are required. In order to further eliminate false positives, the invention also provides a method for enriching ricin in complex samples by using magnetic microspheres. Without knowing the specific active ricin concentration, the gold / quantum dot nanoprobe provided by the present invention can be detected visually by the naked eye through fluorescence quenching (Quenched) and activation (Switch ON).

Description

technical field [0001] The present invention relates to a kind of detection method of natural toxin, be specifically related to a kind of active ricin toxin fast, visual simple detection method, and the gold / quantum dot nano-probe used in this method. Background technique [0002] Ricin is a highly lethal natural toxin, a type II ribosome inactivating protein, with a relative molecular mass of about 64,000Da, composed of two polypeptide chains A and B connected by a disulfide bond, and has Extremely high cytotoxicity (385 times that of VX, 6,000 times that of cyanide, and 10,000 times that of botulinum toxin), which can cause severe poisoning to the body through inhalation, oral administration or intravenous injection. Ricin widely exists in the seeds of the castor plant, and has the characteristics of easy extraction, stable properties, and simple poisoning. At present, there is still no effective means of prevention or treatment for it, so it is frequently used in assassin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6816
CPCC12Q1/6816G01N33/10B82Y5/00G01N33/588G01N2333/42B82Y30/00B82Y20/00B82Y40/00G01N21/6428G01N33/533G01N33/54326G01N33/582G01N33/587G01N2021/6432G01N2021/6439
Inventor 孙洁芳邵兵张晶
Owner BEIJING CENT FOR DISEASE PREVENTION & CONTROL
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