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Polypeptide-metal cluster probe for specifically recognizing CTC membrane protein and application of polypeptide-metal cluster probe for quantitatively detecting expression quantity of membrane protein

A metal cluster and specific technology, applied in the field of medical detection, can solve the problems of inability to enrich CTCs with purity, inability to quantitatively analyze marker proteins, insufficient specificity, etc., and achieve simple preparation steps, less reagents, and low cost. Effect

Pending Publication Date: 2022-03-01
BEIJING UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this process is not specific enough to enrich CTCs with higher purity
At the same time, the current analysis method can only determine the number of CTCs in the blood, but cannot quantitatively analyze the important marker proteins expressed by CTC cells, and the expression of marker proteins is very important for predicting the invasion and metastasis of subsequent tumors. important reference value

Method used

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  • Polypeptide-metal cluster probe for specifically recognizing CTC membrane protein and application of polypeptide-metal cluster probe for quantitatively detecting expression quantity of membrane protein
  • Polypeptide-metal cluster probe for specifically recognizing CTC membrane protein and application of polypeptide-metal cluster probe for quantitatively detecting expression quantity of membrane protein
  • Polypeptide-metal cluster probe for specifically recognizing CTC membrane protein and application of polypeptide-metal cluster probe for quantitatively detecting expression quantity of membrane protein

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preparation example Construction

[0051] The invention provides a preparation method of the polypeptide-metal cluster probe, comprising the following steps:

[0052] 1) drop the metal source solution into the targeting polypeptide solution and mix to obtain a polypeptide-metal source mixed solution;

[0053] 2) performing a reduction reaction on the metal source in the polypeptide-metal source mixture in step 1), and the formed metal atoms form clusters and bind to the targeted polypeptide to obtain polypeptide-metal clusters.

[0054] In the invention, the metal source solution is dropped into the targeted polypeptide solution and mixed to obtain a polypeptide-metal source mixed solution.

[0055] In the present invention, there is no special limitation on the type of the metal source, and metal sources well known in the art can be used. In the embodiment of the present invention, the metal source is HAuCl 4 solution. The concentration of the metal source solution is preferably 20-30 mM, more preferably 25...

Embodiment 1

[0067] (1) In this embodiment, the polypeptide-gold cluster 1 is used, and the targeting peptide is a polypeptide targeting the cell membrane protein N-cadherin, and the sequence is: H 2 N-SWTLYTPSGQSKKKKKGYCC-COOH (Peptide1), the specific preparation steps are as follows:

[0068] Prepare a clean 5mL small glass bottle and a magnet with a diameter of 1cm. Dissolve 2mg of peptide dry powder in 596μL of ultrapure water, slowly add it dropwise into the small glass bottle with the magnet, and stir at 400rpm / min. 3min. 36 μL of 25 mM HAuCl 4 The solution was slowly added dropwise to the above solution and stirred thoroughly for 3 min. Add 179 μL of 0.5M NaOH solution dropwise to the above system, and continue to stir for 5 minutes, then wrap the above system with tinfoil paper and place it on a magnetic stirrer with a set temperature of 37°C, and stir at 600rpm / min to avoid Light reaction 26h. After the reaction is completed and cooled to room temperature, the peptide-gold clu...

Embodiment 2

[0080] (1) Using polypeptide-gold cluster 2, the targeting peptide is a polypeptide targeting cell membrane protein N-cadherin, the sequence is: H 2 N-CCYSWTLYTPSGQSKKKKKG-COOH (Peptide2), the specific preparation steps are as follows:

[0081] Prepare a clean 5mL small glass bottle and a magnet with a diameter of 1cm. Dissolve 2mg of peptide dry powder in 596μL of ultrapure water, slowly add it dropwise into the small glass bottle with the magnet, and stir at 400rpm / min. 3min. 36 μL of 25 mM HAuCl 4 The solution was slowly added dropwise to the above solution and stirred thoroughly for 3 min. Add 358 μL of 0.5M NaOH solution dropwise to the above system, and continue to stir for 5 minutes, then wrap the above system with tinfoil paper and place it on a magnetic stirrer with a set temperature of 37°C, and stir at a speed of 600rpm / min to avoid Light reaction 26h. After the reaction is completed, cool down to room temperature to obtain the targeting peptide-modified polypep...

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Abstract

The invention provides a polypeptide-metal cluster probe for specifically recognizing membrane protein of a circulating tumor cell (CTC) and application of the polypeptide-metal cluster probe in quantitative detection, and belongs to the technical field of medical detection. The polypeptide-modified metal cluster probe prepared by the invention contains a metal cluster core and a target peptide which is specifically combined with target cell membrane protein. The prepared polypeptide-metal cluster probe targets the membrane protein expressed by CTC, and a laser ablation inductively coupled plasma mass spectrometry technology is utilized to perform rapid and accurate in-situ quantitative analysis on the peripheral blood CTC target membrane protein of a clinical tumor patient, so that an important reference is provided for tumor invasion and metastasis behaviors, grading and prognosis of the clinical tumor patient.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a polypeptide-metal cluster probe for specifically recognizing CTC membrane protein and its application in quantitative detection of membrane protein expression. Background technique [0002] Metal clusters are composed of several, dozens or hundreds of metal atoms, usually below 2.0 nanometers, a new type of material with many special physical and chemical properties. Due to their special physical and chemical properties, such as ultra-small size, fluorescence characteristics, catalytic properties, precise structural composition, and good biocompatibility, metal clusters have become a research hotspot in many fields. Based on the unique fluorescent properties and precise structural composition of metal clusters, they have been applied to the detection and quantification of tumor-associated proteins. [0003] Methods for isolating circulating tumor cells (C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K7/08C07K1/13G01N33/68G01N27/626
CPCC07K7/06C07K7/08G01N33/683G01N27/628
Inventor 高靓李函高学云
Owner BEIJING UNIV OF TECH
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