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Hyperpolarization <129>Xe switch type magnetic resonance molecular probe for identifying diamine oxidase

A technology of diamine oxidase and molecular probe, applied in the field of biochemical analysis, can solve the problems of fluorescence quenching effect, weak penetration, unable to obtain the spatial distribution information of diamine oxidase, etc. good effect

Inactive Publication Date: 2018-12-25
WUHAN INST OF PHYSICS & MATHEMATICS CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Fluorescence method has high sensitivity and can be used for qualitative or quantitative analysis, but it is interfered by fluorescence quenching effect and scattered light (Gui Li, Tian Hong, Zheng Jian, et al. Simultaneous determination of four species in mouse brain tissue by HPLC fluorescence method) Amino acid neurotransmitters[D].,2009.)
The coupling enzyme method has high sensitivity and strong specificity, but it is prone to false positives due to the interference of autoantibodies and heterophilic antibodies (Chen Yuanxiang. Evaluation of the diagnostic performance of recombinant Bacillus fastidiosa uricase [D]. Chongqing University of Technology, 2012.)
Not only that, these two methods have weak penetration and cannot obtain the spatial distribution information of diamine oxidase

Method used

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  • Hyperpolarization &lt;129&gt;Xe switch type magnetic resonance molecular probe for identifying diamine oxidase
  • Hyperpolarization &lt;129&gt;Xe switch type magnetic resonance molecular probe for identifying diamine oxidase
  • Hyperpolarization &lt;129&gt;Xe switch type magnetic resonance molecular probe for identifying diamine oxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 The determination of the binding constant of cucurbit [6] urea and putrescine hydrochloride, the steps are as follows:

[0038] (1) Get ammonium acetate and be dissolved in ultrapure water, be configured into the solution that ammonium acetate concentration is 20mM, be recorded as solution 1;

[0039] (2) Take cucurbit [6] urea and dissolve it in solution 1, sonicate until cucurbit [6] urea is completely dissolved, and prepare a solution with a cucurbit [6] urea concentration of 50 μM, which is recorded as solution 2;

[0040] (3) Get putrescine hydrochloride and be dissolved in solution 1, be configured into the solution that putrescine hydrochloride concentration is 1mM, be recorded as solution 3;

[0041] (4) Take solutions 2 and 3, dilute cucurbit[6]urea to a concentration of 22.5 μM and putrescine hydrochloride to a concentration of 0.2 mM with solution 1, respectively, and record them as solutions 4 and 5, respectively;

[0042] (5) Take solutions 4 and...

Embodiment 2

[0046] Embodiment 2 Magnetic resonance instrument 129 The identification of diamine oxidase in Xe spectrum mode, the steps are as follows:

[0047] (1) Get ammonium acetate and be dissolved in ultrapure water, be configured into the solution that ammonium acetate concentration is 20mM, be recorded as solution 1;

[0048] (2) Take cucurbit [6] urea and dissolve it in solution 1, sonicate until cucurbit [6] urea is completely dissolved, and prepare a solution with a cucurbit [6] urea concentration of 50 μM, which is recorded as solution 2;

[0049] (3) Dissolve putrescine hydrochloride in solution 2, and configure a solution with a concentration of putrescine hydrochloride of 100 μM, which is denoted as solution 3;

[0050] (4) Take solution 3, adjust its pH value to 7.2 with acetic acid and ammonia water, and record it as solution 4;

[0051] (5) Take solution 4, add solid diamine oxidase to a final concentration of 2mg / mL, and react on a constant temperature culture shaker (...

Embodiment 3

[0058] Example 3 A hyperpolarization that recognizes diamine oxidase 129 Xe switch-type magnetic resonance molecular probe, which is used in magnetic resonance spectrometer 129 The method for recognizing diamine oxidase in Xe image mode comprises the following steps:

[0059] (1) Get ammonium acetate and be dissolved in ultrapure water, be configured into the solution that ammonium acetate concentration is 20mM, be recorded as solution 1;

[0060] (2) Take cucurbit [6] urea and dissolve it in solution 1, sonicate until cucurbit [6] urea is completely dissolved, and prepare a solution with a cucurbit [6] urea concentration of 50 μM, which is recorded as solution 2;

[0061] (3) Dissolve putrescine hydrochloride in solution 2, and configure a solution with a concentration of putrescine hydrochloride of 100 μM, which is denoted as solution 3;

[0062] (4) Take solution 3, adjust the pH value to 7.2 with acetic acid and ammonia water, and record it as solution 4;

[0063] (5) T...

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Abstract

The invention belongs to the technical field of biochemical analysis and particularly relates to a hyperpolarization <129>Xe switch type magnetic resonance molecular probe for identifying diamine oxidase. The hyperpolarization <129>Xe switch type magnetic resonance molecular probe comprises cucurbit[6]uril and putrescine hydrochloride. The principle of the molecular probe is characterized in thata substrate (putrescine hydrochloride) with high cucurbit[6]uril binding force is converted by diamine oxidase into a product (1-pyrroline) which does not combine with cucurbit[6]uril; before enzymatic reaction, a <129>Xe hyperpolarization chemical exchange saturation transfer signal in the cucurbit[6]uril is inhibited by strong cucurbit[6]uril conjugate until the enzymatic reaction oxidizes the substrate into the product; due to the fact that the binding constant of putrescine hydrochloride@cucurbit[6]uril is several magnitude orders larger than that of <129>Xe@cucurbit[6]uril and (1-pyrroline)@cucurbit[6]uril cannot combine, the <129>Xe hyperpolarization chemical exchange saturation transfer signal in the cucurbit[6]uril appears after the enzymatic reaction through enhanced <129>Xe cucurbit[6]uril interaction, and diamine oxidase identification is achieved through the <129>Xe hyperpolarization chemical exchange saturation transfer signal in the cucurbit[6]uril.

Description

technical field [0001] The invention relates to the technical field of biochemical analysis, in particular to a hyperpolarization method for recognizing diamine oxidase 129 Xe-switched magnetic resonance molecular probes. Background technique [0002] Diamine oxidase is an intracellular enzyme that catalyzes diamine in mammalian small intestinal villous epithelial cells. It can protect intestinal mucosa by regulating intracellular ion balance, affecting conduction pathways, and promoting cell repair (Hou Yue, Gong Ping, Yang Yu, et al. Progress in clinical application of serum diamine oxidase assay [J]. Chinese Experimental Diagnostics, 2015:1985-1987.). In addition, diamine oxidase is closely related to the synthesis of nucleic acid and protein in the small intestinal mucosa (Xi Jiancheng, Ma Yuanzheng, Li Junyou, et al. Changes of diamine oxidase activity in small intestinal tissue during traumatic sepsis[J]. Zhonghua Emergency Clinic Medical Journal, 2005,14(5):421-422....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/26G01N24/08
CPCC12Q1/26G01N24/088G01N2333/90638
Inventor 周欣张斌郭茜旎孙献平刘买利叶朝辉
Owner WUHAN INST OF PHYSICS & MATHEMATICS CHINESE ACADEMY OF SCI
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