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Halloysite coating used for capturing tumour cells and preparation method and application thereof

A technology of tumor cells and halloysite, which is applied in the field of halloysite coating and its preparation, can solve the problems of difficulty in forming rough surface of halloysite, inability to realize large-area preparation, uneven coating, etc., and achieve strong adhesion , low cost and high production efficiency

Active Publication Date: 2016-10-12
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention prepares halloysite into a rough surface by spraying, which can not only effectively overcome the disadvantages of difficulty in forming the rough surface of halloysite by the common direct solution volatilization and drying method, but also overcome the uneven coating caused by coating plastic pipes with solution. The shortcomings of uniformity and poor adhesion can also overcome the shortcomings of traditional nano-surface preparation methods that cannot achieve large-area preparation. It is a new method for manufacturing halloysite nano-rough surfaces that can be used for tumor cell capture.

Method used

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  • Halloysite coating used for capturing tumour cells and preparation method and application thereof
  • Halloysite coating used for capturing tumour cells and preparation method and application thereof
  • Halloysite coating used for capturing tumour cells and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0040]Prepare the dispersion liquid of halloysite, wherein 2 parts of halloysite, 20 parts of deionized water, 80 parts of ethanol, 1 part of γ-aminopropyltriethoxysilane, 0.5 part of surfactant sodium polystyrene sulfonate, The above components are mixed evenly by mechanical stirring at room temperature (400r / min, stirring for 30min), and sprayed on the glass sheet preheated at 100°C with a spray gun; the coating is fully volatilized and dried at 60°C, and then the halloysite coating is soaked into a 5 μg / mL Anti-EpCAM (purchased from Sigma-Aldrich) antibody solution in PBS, removed after 10 min and washed three times with phosphate buffered saline to obtain an antibody-modified halloysite coating.

[0041] attached figure 1 AFM image of the surface of the antibody-modified halloysite coating prepared in Example 1. from figure 1 It can be seen that the surface of the coating is uniform, the thickness is uniform, and the mean square roughness is 52.6nm, which is suitable for...

Embodiment 2

[0044] Prepare the dispersion liquid of halloysite, wherein 5 parts of halloysite, 10 parts of deionized water, 90 parts of ethanol, 2 parts of γ-epoxypropyltriethoxysilane, 0.8 part of surfactant sodium hexametaphosphate, Mix the above components uniformly at room temperature with mechanical stirring (400r / min, stirring for 30min), and spray it on a silicon wafer preheated at 80°C with a spray gun; fully volatilize and dry the coating at 60°C, and then apply the halloysite After the coating was soaked in a PBS solution of 10 μg / mL anti-HER2 (purchased from Sigma-Aldrich) antibody for 30 min, it was washed three times with phosphate buffered saline to obtain an antibody-modified halloysite coating.

[0045] The obtained antibody-modified halloysite coating was used to capture human prostate cancer cells, and the antibody-modified halloysite surface was placed in a cell culture plate, and a blank glass slide was used as a blank control. A measured number of human prostate cance...

Embodiment 3

[0048] Prepare the dispersion liquid of halloysite, wherein 10 parts of halloysite, 100 parts of methanol, 0.5 part of γ-methacryloxypropyl trimethoxysilane, 1.5 parts of surfactant sodium pyrophosphate, the above components in Ultrasonic at 800W for 30 minutes to mix evenly, and then spray it on the aluminum sheet preheated at 50°C with a spray gun; fully volatilize and dry the coating at 80°C, and then soak the halloysite coating into the PBS solution of 30μg / mL anti-HER2 antibody After 20 min, wash with phosphate buffered saline for 3 times to obtain the antibody-modified halloysite coating.

[0049] The resulting antibody-modified halloysite coating was used to capture human liver cancer cells, the surface of the antibody-modified halloysite was placed in a cell culture plate, and a measured number of human liver cancer cells was added to the cell culture medium (DMEM medium, 10% fetal bovine serum), and then the cell culture solution added with a certain amount of cancer ...

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Abstract

The invention belongs to the field of inorganic nanometer materials and biosensors, and discloses a halloysite coating used for capturing tumour cells and a preparation method and application thereof. Specific halloysite nanometer structures are prepared on different substrates through a coating method, and target cell specific recognition molecules are fixed. Or a rough nanometer structure is directly used, the reinforced three-dimensional topology mutual function between the cell surface structure and the substrate rough nanometer structure is used, and more target cell recognition molecules can be fixed on the nanometer rough structure compared with a smooth surface. The flow speed of fluid in blood can be lowered based on the rough nanometer surface structure, and therefore the chance of interacting with cells is increased, the cell capturing efficiency of the halloysite nanometer structure is improved, and the 3h capturing rate for most tumour cells exceeds 80%. The method is easy to operate, low in energy consumption and high in preparation efficiency. In the whole preparation process of the substrates, no toxic and harmful substances are generated, and no pollution is caused to the environment.

Description

technical field [0001] The invention belongs to the field of inorganic nanometer materials and biosensors, in particular to a halloysite coating for capturing tumor cells, a preparation method and application thereof. Background technique [0002] Cancer is one of the major diseases that threaten human health and life in the world. The main reason why cancer is difficult to cure and has a high mortality rate is the existence of cancer metastasis, and the death of more than 90% of cancer patients is directly related to the metastasis of tumor cells. During metastasis, cancer cells from the primary epithelial tumor break off and invade the circulating blood system. As the blood circulates and grows into new tumor tissue (metastasis) in other tissues and organ locations, the cancer cells present in the circulating blood are called circulating tumor cells (Circulating Tumor Cells, CTCs). Detecting the existence and number changes of CTCs in circulating blood is a non-destructi...

Claims

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Application Information

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IPC IPC(8): B05D1/28G01N33/574
CPCB05D1/28G01N33/574
Inventor 刘明贤何瑞周长忍
Owner JINAN UNIVERSITY