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Capture sieve and preparation method thereof

A capture and sieve technology, which is applied in the field of medical devices, can solve the problems of complex capture sieve structure and preparation process, low elution rate, and great impact on subsequent cell processing, and achieves controllable amount of thiol introduction and fast reaction process. , the effect of uniform reaction products

Pending Publication Date: 2020-05-19
上海邦先医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the structure and preparation process of the capture screen are relatively complicated
On the other hand, after specific cell capture in the existing capture screen, the elution of specific cells is relatively complicated or the elution rate is low, which has a great impact on the subsequent cell processing

Method used

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  • Capture sieve and preparation method thereof
  • Capture sieve and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-1

[0055] Example 1-1 Preparation of thiolated sodium alginate

[0056] Step 1: Take 200mg of sodium alginate with a molecular weight of 34KDa and add it to a 250mL single-necked flask, add 120mL of MES buffer (pH=3), and stir to dissolve with a glass rod. Then add 17 mg of 3,3'-dithiobis(hydrazine propionate), and continue stirring until the system is uniform.

[0057] Step 2: Add 50 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide to the single-necked flask, keep the pH of the system at 3, and continue the reaction for 12 hours.

[0058] Step 3: Add 0.5 g of tris(2-carbonylethyl)phosphorus hydrochloride into the single-necked flask, and react for 12 hours.

[0059] Step 4: Take out the solution in the single-necked flask, filter the reacted solution with a filter membrane with a pore size of 0.2 um, and inject the filtered solution into a dialysis bag with a pore size of 30 kDa.

[0060] Step 5: Add 200 mL of 10 mol / L HCl to 500 mL of purified water as a dialysate and add ...

Embodiment 1-2

[0062] Example 1-2 Preparation of thiolated sodium alginate

[0063] Step 1: Take 150mg of sodium alginate with a molecular weight of 20kDa and add it to a 250mL single-necked flask, add 100mL of MES buffer solution (pH=4.5), and stir to dissolve with a glass rod. Then add 16.5 mg of 3,3'-dithiobis(hydrazine propionate), and continue stirring until the system is uniform.

[0064] Step 2: Add 50 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide into the single-necked flask, keep the pH of the system at 4.5, and continue the reaction for 5 hours.

[0065] Step 3: Add 0.05 g of tris(2-carbonylethyl)phosphorus hydrochloride into the single-necked flask, and react for 40 hours.

[0066] Step 4: Take out the solution in the single-necked flask, filter the reacted solution with a filter membrane with a pore size of 5 μm, and inject the filtered solution into a dialysis bag with a pore size of 20 kDa.

[0067] Step 5: Add 100 mL, 23 mol / L HCl into 1000 mL of purified water as a di...

Embodiment 1-3

[0069] Example 1-3 Preparation of thiolated sodium alginate

[0070] Step 1: Take 200mg of sodium alginate with a molecular weight of 34KDa and add it to a 250mL single-necked flask, add 120mL of MES buffer solution (pH=6), and stir to dissolve with a glass rod. Then add 30 mg of 3,3'-dithiobis(hydrazine propionate), and continue stirring until the system is uniform.

[0071] Step 2: Add 120 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide into the single-necked flask, keep the pH of the system at 6, and continue the reaction for 5 hours.

[0072] Step 3: Add 0.5 g of tris(2-carbonylethyl)phosphorus hydrochloride into the single-necked flask, and react for 10 h.

[0073] Step 4: Take out the solution in the single-necked flask, filter the reacted solution with a filter membrane with a pore size of 0.2 um, and inject the filtered solution into a dialysis bag with a pore size of 30 kDa.

[0074] Step 5: Add 100 mL, 1 mol / L HCl into 1000 mL of purified water as a dialysate a...

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Abstract

The invention relates to a capture sieve. The screen mesh comprises a screen mesh framework and captured objects arranged on the screen mesh framework. The capture object can be specifically combinedwith target cells or biomolecules, wherein the screen framework is provided with metal atoms, the screen framework and the captured object are connected through a sulfhydrylation substance, the sulfhydrylation substance and the screen framework react to form a metal-sulfur bond to be combined, and the sulfhydrylation substance and the captured object react to form an amido bond to be combined; wherein the sulfhydrylation substance is one or a combination of two of sulfhydrylation sodium alginate and sulfhydrylation hyaluronic acid. According to the capturing screen, the sulfhydrylation substance is connected to the surface of the screen framework through the metal-sulfur bond. According to the invention, the sulfhydrylated substance is used as a capturing substance, the amido bond is formed between the sulfhydrylated substance and the capturing substance, the capturing substance is connected to the screen skeleton through the sulfhydrylated substance, and the sulfhydrylated sodium alginate or sulfhydrylated hyaluronic acid is adopted, so that the captured substance can be conveniently eluted subsequently on the premise of ensuring higher capturing efficiency.

Description

technical field [0001] The invention belongs to the technical field of medical devices, and in particular relates to a capture sieve connecting capture objects through thiol substances and a preparation method of the capture sieve. Background technique [0002] Microfluidic chip technology integrates the basic operation process of biology, chemistry and other laboratories into a chip, which has the advantages of integration, high throughput, fast detection, convenient operation, small sample size and low energy consumption. It has more and more broad application prospects in many fields of scientific research and life, such as drug research. The means for cell separation and capture on a chip involve many fields such as light, electricity, sound, magnetism, fluid mechanics, mechanical processing and chemical methods. Micromachining technology combined with hydrodynamic control for the capture of whole and single cell and bacterial samples is currently the most effective fix...

Claims

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Application Information

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IPC IPC(8): G01N33/569C12N5/09
CPCG01N33/56966C12N5/0693C12N2509/00
Inventor 颜菁相鸿坤杨天成
Owner 上海邦先医疗科技有限公司
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