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Lipoprotein capillary coating and the preparation method thereof

A lipoprotein and capillary technology, applied in the field of lipoprotein capillary coating and its preparation, can solve the problems of limited practical application, high dependence on the construction of phospholipid double-layer coating structure, etc., and achieve the effect of good biocompatibility

Inactive Publication Date: 2010-06-23
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the construction of a good phospholipid bilayer coating structure is highly dependent on the coating conditions, such as the size and type of phospholipid vesicles formed by self-assembly, which greatly limits the practical application of this method.

Method used

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  • Lipoprotein capillary coating and the preparation method thereof
  • Lipoprotein capillary coating and the preparation method thereof
  • Lipoprotein capillary coating and the preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1 physical adsorption method prepares low-density lipoprotein capillary coating

[0021] Weigh 5.0 mg of low-density lipoprotein freeze-dried powder, fully dissolve in 5.0 mL of 25 mmoL phosphate buffer solution (pH=7.4), and prepare a coating solution of 1.0 mg / mL. Pass the coating solution through the pretreated capillary under 10psi pressure for 15 minutes, then seal both ends of the capillary with silicone rubber, fill the capillary with the above-mentioned low-density lipoprotein coating solution, and let it stand at room temperature for 2-4 hours . Fill the coating solution again, seal both ends of the capillary, and let it stand at room temperature for more than 2 hours. After completing the above operations, rinse the capillary with 25mmoL phosphate buffer solution (pH=7.4) for 30 minutes to remove the low-density lipoprotein not bound to the tube wall, seal both ends of the capillary with silicone rubber, and store in a refrigerator at 4°C.

[0022...

Embodiment 2

[0023] The preparation of the capillary of embodiment 2 aminopropyl modifications

[0024] Pipette 0.1mL of γ-aminopropyltrimethoxysilane, dissolve it in 1.0mL of water (dilute acetic acid to adjust pH 5)-methanol (2:8, volume ratio) mixed solution, mix well and quickly fill into In the pretreated capillary, seal both ends of the capillary with silicon rubber, and react at room temperature for 2-4 hours. After the reaction is completed, the capillary is washed with methanol and water for 30 minutes each, and then the capillary is aged and dried in an oven at 105-120° C. for more than 2 hours under the protection of high-purity nitrogen. Aminopropyl-modified capillaries were obtained through the above-mentioned treatment steps.

Embodiment 3

[0025] Embodiment 3 uses glutaraldehyde as the chemical bonding method of coupling agent to prepare low-density lipoprotein capillary coating

[0026] Pipette 0.5mL of 50% glutaraldehyde aqueous solution, join in 4.5mL of 25mmoL phosphate buffered saline solution (pH=7.4), after thorough mixing, fill in the aminopropyl group prepared by Example 2 under the pressure of 5psi After modifying the capillary, seal both ends of the capillary with silicone rubber after 1 hour, and let it stand at room temperature for 2-6 hours. Then, 25 mmoL phosphate buffer solution (pH=7.4) was used to flush out the unreacted glutaraldehyde solution to obtain a glutaraldehyde-modified capillary.

[0027] Prepare a 1.0 mg / mL low-density lipoprotein coating solution according to the method described in Example 1, fill the solution into the above-mentioned capillary treated with glutaraldehyde, seal both ends after 30 minutes, and continue the reaction at room temperature 2-4 hours. During this react...

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Abstract

The invention relates to a lipoprotein capillary coating and the preparation method thereof. The coating can be prepared with the physisorphtion and chemical bonding methods. In the physisorphtion, a noncovalent interaction between lipoprotein solution with a concentration ranging from 0.01 to 10mg / ml and the internal wall of a capillary is performed and the lipoprotein solution is self-assembled to the internal surface to form the coating. In the chemical bonding method, the capillary is modified with aminopropyl siloxane and the amino is introduced and then the glutaraldehyde or N, N-DSS or 1-ethide-3-(3-dimethyl aminopropyl)-carbodiimide / N-hydroxysuccinimide is used as the resin acceptor to covalently connect the amino on the capillary wall with the amino acid residues on the lipoprotein on the conditions of buffer solution with a pH ranging from 4 to 9 and lipoprotein with a concentration ranging from 0.1 to 10mg / ml for the purpose of obtain the lipoprotein coating. The coating preserves the biological activity, can be used for suppressing the absorption and electroosmotic adjustment on the surface of the capillary and also can be used for biochemical separation analysis.

Description

technical field [0001] The invention relates to a lipoprotein capillary coating and a preparation method thereof, which are suitable for biochemical separation analysis, clinical diagnosis and environmental pollutant toxicity analysis by capillary electrophoresis and electrochromatography. Background technique [0002] Capillary electrophoresis (CE) and capillary electrochromatography (CEC) technologies are the products of the combination of classic electrophoresis technology and modern microcolumn chromatography separation technology. Molecular analysis becomes possible. In recent years, with the rapid development of life sciences and environmental health, capillary electrophoresis and electrochromatography have been widely used in the analysis and identification of biological macromolecules (such as protein separation, DNA fragment and sequence analysis, etc.), clinical disease diagnosis and clinical drug monitoring, Metabolic research, pathological research and other asp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
Inventor 汪海林尹俊发宋玉玲宋茂勇
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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