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Blotting material used for specific recognition of cells, and preparation and applications thereof

A technology of imprinted materials and specificity, which is applied in the direction of analytical materials, instruments, measuring devices, etc., can solve the problems of inability to obtain antibodies, poor cell purity and sensitivity, and inability to perform effectively, so as to achieve good biocompatibility and stability, and improve Purity and Sensitivity, Ease of Handling Effects

Active Publication Date: 2017-05-10
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the uncertainty of the expression of antigens on the cell surface, antibodies against important cells cannot be obtained, so that the technology based on immune principles cannot effectively play its role
In addition, even the same cell has different physical properties under different conditions, so cells obtained based on physical property techniques are often poor in purity and sensitivity (Catherine Alix-Panabières et al, Nat. Rev. Cancer, 2014, 14, 623- 631). Therefore, it is very important to develop a material that can not only replace antibodies, but also specifically recognize and enrich cells

Method used

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  • Blotting material used for specific recognition of cells, and preparation and applications thereof
  • Blotting material used for specific recognition of cells, and preparation and applications thereof
  • Blotting material used for specific recognition of cells, and preparation and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Preparation of cell imprinted material (MIP1) based on SMMC-7721 cell template

[0027] SMMC-7721 cells were cultured on the surface of the twelve-well plate matrix material, when about 80% of the bottom of the culture plate was covered ( figure 1 ), wash the cells with a phosphate buffered saline (PBS) solution of pH=7.2, then add a mass concentration of 4% paraformaldehyde solution to fix the cells, after 5 minutes, wash and remove the paraformaldehyde solution with a PBS solution of pH=7 . Then add 1 ml of pre-polymerized aqueous solution consisting of 30% acrylamide and 1% methylenebisacrylamide to each well of the twelve-well plate, and then add 1 microliter of tetramethylethyl ether. Diamine and 10 microliters of ammonium persulfate aqueous solution with a mass concentration of 10% initiate the curing reaction of the prepolymerization solution. After 20 minutes, the imprinted layer formed by solidification was separated from the matrix material and the imprinted...

Embodiment 2

[0030] Imprinted material (MIP1) for identification and capture of SMMC-7721 cells

[0031] Make two replicates of cells at a concentration of 1 x 10 6 Cells / ml of SMMC-7721 cells were inoculated on the imprinted surface of MIP1 and the surface of NIP respectively. After 2 hours, the cells were attached to the imprinted sites on the surface of MIP1, but there were no SMMC-7721 cells at the non-imprinted sites and the NIP surface. Attach ( Figure 4 ), indicating that the imprinted site of MIP1 can identify and capture target cells rapidly, efficiently and sensitively through multiple interactions with SMMC-7721 cells. SMMC-7721 cells were further added to human blood samples. According to the above method, MIP1 also realized the specific recognition and enrichment of SMMC-7721 cells from blood.

Embodiment 3

[0033] Preparation of cell-imprinted material (MIP2) based on MCF-7 cell template

[0034] Same as Example 1, using MCF-7 cells as a template to prepare imprinted material (MIP2) that can specifically recognize and capture MCF-7 cells, such as Figure 5 shown.

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Abstract

The invention relates to a blotting material used for specific recognition of cells, and preparation and applications thereof. According to the blotting material, cells are taken as a template, cell culturing and cell immobilization technology are adopted to immobilize the cells on the surface of a substrate material, curing of a pre-polymerization system is carried out on the surface of the substrate material, the substrate material is removed via separation, and the template cells on the surface of an obtained cured polymer are removed so as to obtain a finished product. The blotting material is high in selectivity and stability, and can be used in identification, capturing, and enriching of cells.

Description

technical field [0001] The invention belongs to functional biological material, its preparation technology and its application in cell identification, capture, release and enrichment, specifically a novel imprinted material which can specifically identify cells, and its preparation and application. Background technique [0002] High-throughput and high-sensitivity detection of cells plays a very important role in tumor diagnosis and treatment, environmental detection and biosensor research (Aherne A. et al., J.Am.Chem.Soc., 1996,118,8771- 8772). At present, the detection of cells is very difficult in an environment with low cell concentration, mainly because the cell content is small and cannot be obtained. Therefore, the premise of realizing cell detection in low cell concentration environment is to be able to identify and enrich the target from the environment. [0003] Cell identification and enrichment Cell identification and enrichment technology is a technology that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J9/26C08F220/56C08F222/38C08G18/76C08G18/32G01N33/569
CPCC08F220/56C08G18/3215C08G18/7671C08J9/26C08J2201/0462C08J2333/26C08J2375/04C08J2381/06G01N33/569C08F222/385
Inventor 张丽华刘路宽杨开广李森武张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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