Difunctional nanoprobe based on lanthanide metal as well as preparation method and application of difunctional nanoprobe

A technology of nano-probes and lanthanide metals, applied in the field of analytical chemistry, can solve the single signal response performance of probes, cannot realize real-time, in-situ, and visualized analysis of biomolecules, and cannot simultaneously realize on-site dynamic visualization of biomolecules and high Sensitive quantitative analysis and other issues to achieve the effect of avoiding false positive signals, realizing quantitative detection, and good response performance

Active Publication Date: 2016-01-20
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Description
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  • Application Information

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Problems solved by technology

However, the detection effect of biomolecules can only be learned through the final mass spectrometry characterization, and real-time, in situ, and visualized analysis of biomolecules cannot be realized
[0004] Traditional probes only have a single signal response performance, so dynamic visualization and high-sensitivity quantitative analysis of biomolecular sites cannot be realized at the same time

Method used

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  • Difunctional nanoprobe based on lanthanide metal as well as preparation method and application of difunctional nanoprobe
  • Difunctional nanoprobe based on lanthanide metal as well as preparation method and application of difunctional nanoprobe
  • Difunctional nanoprobe based on lanthanide metal as well as preparation method and application of difunctional nanoprobe

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Experimental program
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Embodiment 1

[0063] The experimental procedure for preparing nanoprobes by using Eu-BCTOT-labeled peptides to modify gold nanoparticles is as follows: figure 1 shown.

[0064] 1) Optimization of Eu and BCTOT chelation conditions: a certain amount of EuCl 3 After being dissolved in deionized water, it was added to the BCTOT solution, and after incubation at 50°C for a period of time, the ultraviolet and fluorescence spectra of the system were measured. Through spectral analysis of the solution obtained after Eu and BCTOT chelation, the maximum excitation and emission wavelengths of Eu-BCTOT chelate are 350nm and 614nm respectively, which is the characteristic emission spectrum of Eu, indicating that Eu and BCTOT are successfully chelated. And BCTOT can indeed play a very good sensitization effect on the luminescence of Eu.

[0065] Then, the optimal chelation ratio of Eu to BCTOT was investigated, and the results showed that when the molar ratio of Eu to BCTOT was in the range of 0.1-10:1...

Embodiment 2

[0077] Example 2. Using the nanoprobe prepared in Example 1 to detect exogenous caspase-3 by spectrum and mass spectrometry.

[0078] 1) Different concentrations of exogenous caspase-3 and 10 μM probe were incubated in PBS (pH 7.4) buffer at 37° C. for 2 h. After the reaction, the solution was centrifuged at 120,000 rpm for 10 min, and the supernatant was collected for fluorescence detection. The excitation and emission wavelengths detected were 350 nm and 614 nm, respectively. The result is as Figure 4 shown.

[0079] Depend on Figure 4It can be seen that as the concentration of caspase-3 increases from 5ng / mL to 70ng / mL, the fluorescence intensity in the reaction system also increases. It shows that the probe of the present invention can be used for the detection of caspase-3; at the same time, it shows that based on the principle of FRET, an open nanoprobe is successfully constructed for the detection of caspase-3.

[0080] At the same time, in order to verify that t...

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Abstract

The invention discloses a difunctional nanoprobe based on lanthanide metal as well as a preparation method and an application of the difunctional nanoprobe. The preparation method of the difunctional nanoprobe based on the lanthanide metal comprises steps as follows: 1), polypeptides containing DEVD amino acid residues are taken as a bridging agent and firstly react with antenna molecules BCTOT (10-bis(5'-chlorosulfo-thiophene-2' -yl)-4,4,5,5,6,6,7,7-octafluorodecane-1,3,8,10-tetraone), and BCTOT modified polypeptides are obtained; 2), the BCTOT modified polypeptides have a chelation reaction with lanthanide metal ions, and a chelated polypeptide solution is obtained; 3), the chelated polypeptide solution reacts with a nanomaterial solution, and the difunctional nanoprobe based on the lanthanide metal is obtained. The difunctional nanoprobe based on the lanthanide metal is applied to caspase-3 spectrum detection, mass spectrometric detection, cell imaging and/or ICP-MS (inductively coupled plasma mass spectrometry) quantitative detection. Dynamic visualization of biomolecule sites and high-sensitivity quantitative analysis can be achieved.

Description

technical field [0001] The invention relates to a bifunctional nanoprobe based on lanthanide metals, a preparation method and application thereof, and belongs to the field of analytical chemistry. Background technique [0002] Dynamic visualization and highly sensitive quantitative analysis of important regulatory molecules in living systems are of great significance to biological and clinical research. Due to the high sensitivity and spatiotemporal resolution of fluorescence detection methods, fluorescence imaging methods have been widely developed in the real-time visual detection of biomolecules and biological processes. However, the quantitative detection of biomolecules using fluorescence methods still faces great challenges. This is because: firstly, ratiometric fluorescent probes can be designed to achieve quantitative detection of proteases using fluorescence resonance energy transfer, but this method requires that the emission spectrum of the donor fluorescence ove...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/25G01N21/75B82Y30/00B82Y40/00
Inventor 钱小红秦伟捷冯端
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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