Rare-earth cage-like coordination compound metal fluorescent dual-mode label for flow cytometry and preparation method of rare-earth cage-like coordination compound metal fluorescent dual-mode label

A flow cytometry and complex technology, applied in the field of rare earth cage complex metal fluorescent dual-mode label and preparation, can solve the problem of difficulty in meeting flow cytometry detection requirements, weak rare earth metal fluorescence ability, high price, etc. problem, to achieve the effect of reducing leakage, improving fluorescence intensity and signal strength

Active Publication Date: 2021-05-11
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the one hand, the labels on the market are usually imported from abroad and are expensive. The price of a standard kit (containing 40*100 micrograms of rare earth coordination polymers, which can carry out 40 labeling experiments) is usually more than 50,000 RMB
On the other hand, the metal signals on the labels on the market can be effectively detected by mass spectrometry, so as to achieve the effect of single-cell detection. However, the fluorescence ability of rare earth metals is weak, which cannot be well applied to traditional flow cytometry detection.
However, the fluorescence / metal detection reagents reported in the existing literature have relatively weak fluorescence signal and metal signal intensity, which is difficult to meet the detection requirements of flow cytometry and mass spectrometry flow cytometry.

Method used

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  • Rare-earth cage-like coordination compound metal fluorescent dual-mode label for flow cytometry and preparation method of rare-earth cage-like coordination compound metal fluorescent dual-mode label
  • Rare-earth cage-like coordination compound metal fluorescent dual-mode label for flow cytometry and preparation method of rare-earth cage-like coordination compound metal fluorescent dual-mode label
  • Rare-earth cage-like coordination compound metal fluorescent dual-mode label for flow cytometry and preparation method of rare-earth cage-like coordination compound metal fluorescent dual-mode label

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Effect test

preparation example Construction

[0056] The preparation method of the rare earth cage ligand metal fluorescent dual-mode label for flow cytometry comprises the following steps:

[0057] 1) Prepare block monodisperse copolymer, its structure is as shown in formula (Ⅰ-5);

[0058] The block monodisperse copolymer is prepared by N-hydroxysuccinimide acrylate, N,N-dimethylacrylamide, tert-butylbenzocarbodisulfate and azobisisovaleronitrile. The synthetic route of block monodisperse copolymer is:

[0059]

[0060] Among them, formula (I-1) is N-hydroxysuccinimide acrylate, formula (I-2) is N,N-dimethylacrylamide, formula (I-3) is tert-butylbenzocarbodi Sulfate ester, formula (I-4) is azobisisovaleronitrile.

[0061] 2) Prepared by the block monodisperse copolymer, rare earth cage complex, 1,8-bis(maleimide)-3,6-dioxa-octane as shown in formula (I-8) The polymer with rare earth cage complex is shown, the structure of rare earth cage complex is shown in formula (I-6), 1,8-bis(maleimide)-3,6-bis The structure ...

Embodiment 1

[0071] In this example, the monoclonal antibody is selected as CD-4, Ln 3+ Choose Eu 3+ .

[0072] In this embodiment, the preparation method of the rare earth cage ligand metal fluorescent dual-mode label for flow cytometry includes the following steps:

[0073] 1. Preparation of block monodisperse copolymers

[0074] 1-1) Add 2.46g of N-hydroxysuccinimide acrylate, 0.96g of N,N-dimethylacrylamide, 54mg of tert-butylbenzocarbodisulfate and 16mg of azobisisovaleronitrile to 45mL of dry DMF;

[0075] 1-2) The reaction system was deoxygenated by freezing with liquid nitrogen, and then stirred at 60°C for 16 hours under the protection of nitrogen;

[0076] 1-3) Cool to room temperature, add to 400ML diethyl ether for precipitation, redissolve the obtained solid in 1,4-dioxane, and again precipitate with diethyl ether;

[0077] 1-4) The fixed product obtained after filtration was dried to obtain 3.0 g of the block monodisperse copolymer.

[0078] refer to figure 2 , for bl...

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Abstract

The invention discloses a rare-earth cage-like coordination compound metal fluorescent dual-mode label for flow cytometry, which is obtained by coupling a polymer with a rare-earth cage-like coordination compound as shown in the a formula (I-8) defined in the description with a monoclonal antibody, wherein Ln<3+> is a lanthanide rare earth metal ion. Different from an existing mass spectrum flow type polymer metal chelating label, a DOTA coordination group is ingeniously replaced with a cage-shaped coordination group, on the one hand, the coordination capacity of rare earth ions is improved, leakage of the rare earth ions can be reduced, and signals of the metal label in a flow type mass spectrum are stronger; on the other hand, due to the antenna effect of the cage-like coordination compound, the fluorescence intensity of the rare-earth coordination compound is greatly improved, and the rare-earth coordination compound can be detected by a flow cytometry. According to the invention, the rare earth cage-like coordination compound metal fluorescent dual-mode label for flow cytometry can be prepared at a lower cost, so that the use cost of a mass spectrum flow detection reagent can be greatly reduced.

Description

technical field [0001] The invention relates to the technical field of biological imaging, in particular to a rare earth cage ligand metal fluorescent dual-mode label for flow cytometry and a preparation method. Background technique [0002] Mass cytometry is a new type of single-cell detection technology developed on the basis of flow cytometry. It stains cells with antibodies labeled with metal elements, and then detects metal signals through inductively coupled plasma mass spectrometry. To achieve the purpose of identifying cells. Simultaneous biomarker detection and single-cell analysis by flow cytometry and mass cytometry are of great significance for pathology research, drug release, and signaling network research. The roles of flow cytometry and mass cytometry are complementary. Mass cytometry can provide multiple detection, while flow cytometry has higher sensitivity and detection capacity, and cells will not be detected during the detection process. destroyed. Pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F293/00C08F220/34C08F220/54C08F8/42C08F8/30G09F3/00C07K16/28
CPCC08F293/00C08F220/34C08F220/54C08F8/42C08F8/30G09F3/00C07K16/2812
Inventor 刘志周何良王彤南雪燕白鹏利王辉胡玮
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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