Quantum dot targeting probe kit for detecting tumor of colon cancer

A technology of quantum dots and kits, applied in the field of quantum dot targeted probe kits, can solve problems such as poor photostability, easy photobleaching, and inability to display lesions.

Inactive Publication Date: 2017-04-19
CHANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many examination methods for colon cancer, among which colonoscopy and barium enema examination are commonly used examination methods, but these examinations can only show the lesions inside the colon cavity, but cannot show the lesions outside the cavity, and the detection has a certain degree of pain
[0003] The detection of tumor cells requires a probe that can specifically recognize tumor markers. Traditional organic fluorescent dyes are commonly used probes for biomarkers and cell imaging, but they also have some inherent defects, such as poor photostability, easy photobleaching, emission Spectral peak tailing, etc., which limit its application

Method used

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  • Quantum dot targeting probe kit for detecting tumor of colon cancer
  • Quantum dot targeting probe kit for detecting tumor of colon cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1 Composition and configuration of a quantum dot targeting probe kit for detecting colon cancer tumors:

[0021] The kit includes a) TCP-1-H6 polypeptide, b) TP-1-H6 polypeptide, c) water-soluble quantum dot, d) FITC, e) DAPI.

[0022] A). The polypeptide sequence of TCP-1-H6 is cyclo(1,9)-CTPSPFSHCGP 9 G 2 h 6 .

[0023] B). The sequence of TP-1-H6 polypeptide is TPSPFSHGP 9 G 2 h 6 . Both peptides were synthesized by solid phase synthesis.

[0024] C). The water-soluble quantum dots are quantum dots containing Zn, such as CdSe / ZnS, CdTe / ZnS, CdSe / ZnSe or CdTe / ZnSe. In the quantum dots, glutathione is used as a modifier to convert fat-soluble quantum dots into water-soluble quantum dots.

[0025] D). Fluorescein isothiocyanate (FITC)

[0026] E).4,6-diamidino-2-phenylindole (DAPI)

Embodiment 2

[0027] Embodiment 2 detects Colon26 cells with a quantum dot targeting probe kit for detecting colon cancer tumors, and the process is as follows:

[0028] 1. Mix the peptide and quantum dots at a ratio of 18:1, shake for 10 minutes, and remove excess peptide molecules by ultrafiltration to obtain a quantum dot-polypeptide fluorescent probe.

[0029] 2. Colon 26 cells whose nuclei were stained with DAPI were cultured in RPMI-1640 (Invitrogen) medium and incubated overnight in a carbon dioxide incubator (5% CO2, 37°C).

[0030] 3. After TCP-1-H6-QD was mixed, it was incubated at 37°C for 4 hours, and the unlabeled peptide quantum dots and TP-1-H6-QD were used as a control.

[0031] 4. The incubated cells were washed three times with PBS buffer (pH 7.4).

[0032] 5. Observe the fluorescence imaging under a Nikon C1si laser confocal fluorescence microscope (Nikon, Japan) (excitation wavelength is 488nm).

[0033] The experimental results confirmed that the fluorescence of quant...

Embodiment 3

[0034] Example 3 Using a quantum dot targeting probe kit for detecting colon cancer tumors to detect mouse colon cancer tumor tissue sections, the process is as follows:

[0035] 1. Mix the polypeptide and quantum dots at a ratio of 18:1, shake for 10 minutes, and remove excess polypeptide molecules by ultrafiltration to obtain the quantum dot-polypeptide fluorescent probe.

[0036] 2. Select colon cancer tumor tissue slices from mice containing colon cancer tumors, and select normal colon tissue and brain tissue as controls.

[0037] 3. The blood vessels in the selected tissue were stained with FITC-labeled secondary antibody, and the nuclei were stained with DAPI.

[0038] 4. After mixing tumor tissue or other tissues with TCP-1-H6-QD (100nM), incubate at 37°C for 1h.

[0039] 5. The incubated tissue was washed three times with PBS buffer (pH 7.4).

[0040] 6. Observe the fluorescence imaging under a Nikon C1si laser confocal fluorescence microscope (Nikon, Japan) (excitat...

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Abstract

The invention discloses a quantum dot targeting probe kit for detecting the tumor of colon cancer. The kit comprises (a) TCP-1-H6 polypeptide, (b) TP-1-H6 polypeptide, (c) water soluble quantum dots, (d) FITC, and (e) DAPI. Water soluble quantum dots and targeting TCP-1-H6 polypeptide are coupled through His-tag and taken as a fluorescence probe (TCP-1-H6-QD). TCP-1-H6-QD can specifically label the acceptor of colon 26 cell and tumor tissue slice of colon cancer, and will not recognize normal colon tissues and brain tissues. The provided kit has an important meaning for the diagnosis and treatment of color cancer.

Description

technical field [0001] The invention belongs to the field of nanobiology technology, and relates to a colon cancer tumor detection kit, in particular to a quantum dot targeting probe kit for detecting colon cancer tumor cells and tumor tissues. Background technique [0002] Colon cancer is a common malignant tumor of the digestive tract. Due to the lack of obvious early symptoms and lack of specificity, the symptoms are hidden and difficult to detect when a tumor occurs. There are many examination methods for colon cancer, among which colonoscopy and barium enema examination are commonly used examination methods, but these examinations can only show the lesions inside the colon cavity, but cannot show the lesions outside the cavity, and the detection has a certain degree of pain. [0003] The detection of tumor cells requires a probe that can specifically recognize tumor markers. Traditional organic fluorescent dyes are commonly used probes for biomarkers and cell imaging, b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 王建浩邱琳蒋鹏举柳丽丁淑敏张晨澄滕一万吴昊董冰玉
Owner CHANGZHOU UNIV
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