Nucleotide aptamer-based metal probe as well as preparation method and application thereof

A technology of metal probes and aptamers, which can be used in the direction of material inspection products, measuring devices, instruments, etc., can solve the problems of Aptamer targeting limitations, etc., and achieve the effect of low price, small molecular weight, and good imaging effect

Pending Publication Date: 2021-06-29
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to target PSMA, the Aptamer screened from the synthetic RNA aptamer library in the early stage consists of A10 and A9 consisting of 79 nucleotides, but the targeting of long-sequence high-molecular-weight Aptamers is limited.

Method used

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  • Nucleotide aptamer-based metal probe as well as preparation method and application thereof
  • Nucleotide aptamer-based metal probe as well as preparation method and application thereof
  • Nucleotide aptamer-based metal probe as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Principle of Nucleotide Aptamer Recognition Target Molecule

[0039] Nucleotide aptamers (Aptamers) are usually single-stranded DNA or single-stranded DNA that can bind to target antigens with high affinity and specificity produced in vitro through a process called ligand system evolution (SELEX). RNA oligonucleotides, ranging in length from 15 to 100 nucleotides. Such as figure 1 As shown, the nucleotide aptamer Aptamer recognizes the target molecule, and the combination of the two is an affinity binding process, and the binding body has a three-dimensional structure. The nucleotide sequence of the nucleotide aptamer Aptamer determines its three-dimensional structure that specifically binds to the target molecule. In addition, it can specifically recognize inorganic ions, small organic molecules, peptides, proteins, bacteria, viruses and cells.

Embodiment 2

[0040] Embodiment 2 The synthetic route of the metal probe based on nucleotide aptamer

[0041] Metal-labeled Aptamer probes 167 Er-A10-3.2 by 167 Er metal cations, bifunctional polymers, and -SHC 6 -A10-3.2 Nucleotide aptamer composition. First, triaminepentaacetic acid (DTPA) groups at the first end of the bifunctional polymer chelate 167 Er cations formed polymer-based lanthanide metal-chelating polymers (MCPs ( 167 Er)), where the number of metal ions is determined by the degree of polymer (DP) of the polymer. The DP of MCPs(Ln) polymer is generally 20-25. MCPs ( 167 Er) The maleimide functional group at the second end of the polymer and -SHC 6 The -SH of -A10-3.2 undergoes a Michael addition reaction, and a metal-labeled aptamer probe is synthesized.

[0042] Such as figure 2 As shown, the steps of the metal probe synthesis route based on nucleotide aptamers are:

[0043] Step 1 will 167 Er cations and bifunctional polymers were added to the ultrafiltration tu...

Embodiment 3

[0046] Embodiment 3 Ability of the specific recognition target of the metal probe based on nucleic acid aptamer

[0047] The transmembrane glycoprotein PSMA protein of epithelial cells in prostate cancer PaC tumor tissue slices was selected as the research object, and MCPs ( 167 Er) For the metal label, select Anti-PSMA-SHC 6 -A10-3.2 aptamer Aptamer and Anti-PSMA YPSMA-1 antibody as targeting groups, MCPs ( 167 Er) modified Ab and Aptamer prepared 167 Er-YPSMA-1 and 167 Er-A10-3.2 Two different types of probes. The successful construction of nanoprobes was verified by ultraviolet-visible absorption spectroscopy (UV-Vis) and transmission electron microscopy (TEM). Through traditional immunofluorescence imaging (Immunofluorescence imaging, IF) technology combined with IMC technology, the expression levels of PSMA in selected PaC tumor tissues and normal tissues were compared and analyzed; 167 Er-A10-3.2 and 167 The targeting ability of the Er-YPSMA-1 probe nanoprobe was v...

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Abstract

The invention discloses a nucleotide aptamer-based metal probe as well as a preparation method and application thereof, and relates to the technical field of biological detection. The probe is composed of a 167Er metal cation, a bifunctional polymer and a -SHC6-A10-3. 2 aptamer. The preparation method of the probe comprises the steps of chelating 167Er cations with diethylenetriamine pentaacetic acid (DTPA) in a bifunctional polymer to form a polymer-based 167Er lanthanide metal chelate MCPs (167Er), and carrying out an addition reaction on the successfully chelated MCPs (167Er) and the -SHC6-A10-3.2 nucleic acid aptamer to generate the IMC probe based on the nucleotide aptamer. The metal probe can be used as a non-invasive diagnostic imaging reagent for identifying a target substance, and is used for quantitative and positioning analysis of protein of a cell line or tissue. The nucleotide aptamer-based metal probe disclosed by the invention has the advantages of high sensitivity, accurate positioning, high dyeing accuracy and the like.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a metal probe based on a nucleic acid aptamer and its preparation method and application. Background technique [0002] Single-cell mass spectrometry imaging (IMC) mainly uses lanthanide metal isotope-labeled antibodies (Antibody, Ab) and uses mass spectrometry instead of spectrum as the detection method. It has ultra-low background, no interference between channels, and multi-parameter detection. It is completely different from the traditional mode. advantage features. IMC technology has the advantages of convenient application, diverse labeling targets, and high throughput, and plays an important role in the fields of disease diagnosis and treatment, drug development, and basic biological research. Compared with previous tissue analysis methods, multi-channel IMC imaging technology can provide a comprehensive understanding of tissue composition and biomarker conte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/68G01N27/62
CPCG01N33/57434G01N33/57492G01N33/68G01N27/62
Inventor 丁显廷余友谊党婧琪
Owner SHANGHAI JIAO TONG UNIV
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