Capture sieve and device for capturing cells or biomolecules in solutions

A technology of biomolecules and capture objects, which is applied in the field of devices for capturing biomolecules in solution, and can solve the problems of inability to capture living cells, difficulties in industrialization, and poor specificity

Pending Publication Date: 2017-11-10
HEMOSMART MEDICAL TECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally, there are only 1-10 circulating tumor cells in 10ml blood of cancer patients, and their slow capture speed or poor specificity is an urgent problem to be solved for rapid detection of patient blood samples
[0003] The current sorting methods such as the traditional immunomagnetic bead sorting method have good repeatability, high sensitivity, good specificity, and can quantitatively analyze circulating tumor cells, but the operation speed is slow and the throughput is small
Although membrane microfiltration technology and gradient density centrifugation method are simple to operate and have good cell viability after separation, they have low specificity and high false positive rate
Microfluidic technology is simple to operate and requir...

Method used

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  • Capture sieve and device for capturing cells or biomolecules in solutions
  • Capture sieve and device for capturing cells or biomolecules in solutions
  • Capture sieve and device for capturing cells or biomolecules in solutions

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0065] Herein, a preparation method of the capture sieve for sorting CTCs is given. The preparation method includes:

[0066] (I) selection of sieves;

[0067] Gold sieve: Choose a gold sieve with pores (eg, 64 µm or 40 µm) to maximize contact time with tumor cells while preventing the risk of clogging. The size of the gold sieve used in the following tests is 2×2mm 2 . Larger sizes are preferred depending on the specific operation.

[0068] Stainless steel and gold sieves: 51µm pores are selected and AuPd is coated by magnetron sputtering. Such sieves are cheaper and mechanically stronger than gold, so they are more suitable for integration in devices.

[0069] (II) Pre-functionalization;

[0070] Various cleaning methods were used to prepare the sieves prior to functionalization, including autoclaving, oxygen plasma cleaning, and ultrasonic cleaning in various solutions, including piranha solutions. For example, the best results for a 64 µm gold sieve included sonicat...

example 2

[0080] Experiments were performed to demonstrate the efficiency of the screen in capturing EpCAM expressed by tumor cells. In this case, the mesh opening is 51 μm.

[0081] Cell Selection - Cells with high expression levels of EpCAM protein (such as CaCo2 and MCF7 cells) were used.

[0082] Cell Growth - Grow in DMEM buffer at 37°C.

[0083] Cell preparation and incubation of screens - CaCo2 and MCF7 cells differentiated 1:2. The test was performed on a rotating 37°C hot plate. After isolation, cells were diluted 1:10 in DMEM buffer, and 0.5 ml was used to incubate the capture screen for 1 hour.

[0084] Washing of non-specifically bound cells, the sieves were rinsed with pure water, incubated in pure aqueous solution for 2 minutes, and then rinsed again before microscopic observation.

[0085] Microscopic evaluation of trapped cells: photograph of the sieve taken by the microscope ( Figure 5a and 5b ). Figure 5a CaCo2 cells expressing the target capture molecule capt...

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PUM

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Abstract

The invention discloses a capture sieve and a device for capturing cells or biomolecules in solutions. The capture sieve comprises a net-shaped base body and a capture layer formed on the net-shaped base body, wherein the capture layer comprises capture matter which can be in specific binding with target cells or biomolecules. The capture sieve and the device provided by the invention have high specificity and high flux, are suitable for capturing the cells or capturing the biomolecules in the solutions by adopting the molecules expressed by the cells, and are particularly suitable for capturing and sorting circulating tumor cells.

Description

technical field [0001] The present invention relates to a device for capturing a cell or capturing a biomolecule in solution by a molecule expressed by the cell, in particular to a device for capturing a target cell (eg, circulating tumor cells) or capturing a biomolecule expressed by a target cell A capture screen for target biomolecules in solution and a device having such a capture screen. Background technique [0002] Circulating tumor cells, that is, tumor cells circulating in the blood, are considered to have a significant relationship with distant metastasis of tumors and other issues. Generally, there are only 1-10 circulating tumor cells in 10ml blood of cancer patients, and their slow capture speed or poor specificity is an urgent problem to be solved for rapid detection of patient blood samples. [0003] Current sorting methods such as the traditional immunomagnetic bead sorting method have good repeatability, high sensitivity, and good specificity, and can quant...

Claims

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Application Information

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IPC IPC(8): C12M1/12C12M1/34
CPCC12M41/36C12M47/04
Inventor 查梅特·杰罗姆颜菁鲍尔·沃尔夫冈安德烈斯·克里斯蒂安余子夷杨洋
Owner HEMOSMART MEDICAL TECH LTD
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