Polyatomic element labeling probe and preparation method and application thereof

A technology for labeling probes and atomic elements, applied in biochemical equipment and methods, material inspection products, instruments, etc., can solve the problems of water solubility and stability attenuation, difficulty in controlling the polymerization degree of polymer chains, and difficulty in synthesizing element tags, etc. problem, to achieve the effect of simple purification process

Active Publication Date: 2019-05-31
TSINGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are some inherent problems in the organic polymer chain. On the one hand, its water solubility and stability decrease rapidly with the increase of the chain, which makes it difficult to further increase the number of labels. On the other hand, the polymer chain’s The degree of polymerization is difficult to control, and it is difficult to synthesize element tags with different degrees of polymerization, so it is impossible to effectively adjust the detection linear range and the number of tags according to the sample requirements

Method used

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  • Polyatomic element labeling probe and preparation method and application thereof
  • Polyatomic element labeling probe and preparation method and application thereof
  • Polyatomic element labeling probe and preparation method and application thereof

Examples

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Embodiment 1

[0039]A 180bp long polyalkyne DNA backbone structure was synthesized by PCR, and the polyalkyne DNA was coupled to a macrocyclic metal ion complex containing azide group and Eu modification by click reaction to obtain polyatomic element-labeled probes.

[0040] Its synthesis process is as follows:

[0041] (1) Weigh the macrocyclic metal ion complex DOTA-N containing azide group and Eu modification 3 (Azido-mono-amide-DOTA, purchased from Macrocyclics (Dallas TX): B288) was dissolved in PBS buffer (pH=5.8), prepared into a 30mg / ml mother solution, and stored at 4°C;

[0042] (2) Take another 200 μL centrifuge tube, add DNA polymerase, DNA template, PCR buffer, A, G, C three conventional nucleotides and 5-ethynyl-dUTP, upstream and downstream primers (primer sequences are shown in Table 2 shown), magnesium sulfate, and add deionized water to make up to 50 μL. The DNA polymerase and the corresponding PCR buffer used are matching commercial polymerase and buffer (NewEngland Bio...

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Abstract

The invention discloses a polyatomic element labeling probe and a preparation method and application thereof. The method comprises the steps: synthesizing a DNA molecule containing multiple active groups rapidly by using the PCR technology, taking the DNA molecule as a skeleton structure, coupling a metal macrocyclic compound into the skeleton structure through an orthogonal reaction, and efficiently preparing a multi-labeled element label. The element label is used for traditional immunization, the sensitivity of the element label reaches pg/ml, and the element label is suitable for the fieldof trace analysis and detection of biomacromolecules.

Description

technical field [0001] The invention belongs to the field of biological detection, and relates to a polyatomic element labeling probe, a preparation method and application thereof. Background technique [0002] Element labeling refers to the use of atoms or ions as labels (the "atoms or ions" here can be metal ion macrocyclic compounds or metal atoms, etc.), labeling biomacromolecules, and through biomolecular interactions (antigen-antibody interaction) , DNA hybridization, etc.), inductively coupled plasma (ICP-MS) technology can be used to capture atomic or ion signals, and then directly or indirectly reflect the relationship between the amount of biomolecules. ICP-MS demonstrates the unique advantages of multi-channel analysis and absolute quantitative detection in clinical and life analysis by combining elemental labeling of biomacromolecules, and opens up a new way for quantitative bioanalysis. Among them, almost all isotopes of more than 100 ICP-MS identifiable elemen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/534G01N33/60G01N27/62G01N21/64G01N21/31C12Q1/6816G01N33/574
Inventor 张新荣胡志安孙公伟张钰清邢志张四纯
Owner TSINGHUA UNIV
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