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A kind of anti-protein adsorption capillary column and preparation method thereof

An anti-protein adsorption and capillary column technology, which is applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problems of complex preparation process, high toxicity in the production process, harsh operating conditions, etc., and achieve simple and fast preparation process, excellent anti-protein The effect of adsorption performance

Active Publication Date: 2020-12-22
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] For this reason, the technical problem to be solved by the present invention is that the existing preparation process is complex, the operating conditions are harsh, the production process is highly toxic, the production efficiency is low, the production cost is high, and the prepared anti-protein adsorption capillary electrophoresis covalently bonded coating column In order to solve the problem of poor quality, an anti-protein adsorption capillary column and its preparation method are provided. The process is simple, the production process is environmentally friendly, the production efficiency is high, and the production cost is low. good column quality

Method used

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  • A kind of anti-protein adsorption capillary column and preparation method thereof
  • A kind of anti-protein adsorption capillary column and preparation method thereof
  • A kind of anti-protein adsorption capillary column and preparation method thereof

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preparation example Construction

[0028] The preparation method of the above-mentioned anti-protein adsorption capillary column uses diazo resin and carboxylated chitosan to self-assemble layer by layer on the inner wall of the quartz capillary column through electrostatic interaction, and undergoes a photocuring cross-linking reaction under ultraviolet light to make the original hydrogen The bonds are converted to covalent bonds, thereby forming a stable covalently bonded coating on the inner wall of the quartz capillary. Specifically, the preparation method comprises the following steps:

[0029] S1, the concentration is 0.05-500mg·ml -1 Photosensitive diazo resin solution, water, concentration 0.05-500mg·ml -1 The carboxylated chitosan aqueous solution and water are slowly injected into the capillary successively; the mass ratio of the photosensitive diazo resin to the carboxylated chitosan aqueous solution is 1:1.

[0030] The concentrations of the above-mentioned diazo resin and carboxylated chitosan aq...

Embodiment 1

[0046] This implementation provides a preparation method for an anti-protein adsorption capillary column, including the following steps:

[0047] S0, capillary internal activation pretreatment

[0048] successively with a concentration of 0.1mol·L -1 Sodium hydroxide solution, distilled water, the concentration is 0.1mol L -1 Flush the capillary with hydrochloric acid solution, distilled water and methanol for 30min, 10min, 30min, 10min and 10min respectively, and then use N 2 The activated capillary can be obtained by drying the capillary.

[0049] S1. At a temperature of 25°C, use a concentration of 0.1 mg·ml -1 The photosensitive diazo resin solution (molecular weight 1000) rinses the capillary column for 10 minutes, and distilled water rinses the capillary column for 2 minutes. Then use a concentration of 0.5mg·ml -1 Carboxylated chitosan aqueous solution (weight-average molecular weight 150000) washes the capillary column for 10 minutes, and then uses distilled water...

Embodiment 2

[0053] This implementation provides a preparation method for an anti-protein adsorption capillary column, including the following steps:

[0054] S0, capillary internal activation pretreatment

[0055] successively with a concentration of 0.09mol·L -1 Sodium hydroxide solution, distilled water, the concentration is 0.11mol L -1 Flush the capillary with hydrochloric acid solution, distilled water and methanol for 35min, 8min, 35min, 8min and 8min respectively, and then use N 2 The activated capillary can be obtained by drying the capillary.

[0056] S1. At a temperature of 25°C, the concentration is 100mg·ml -1 The photosensitive diazo resin solution (weight average molecular weight 5500) washes the capillary column for 10 minutes, distilled water washes the capillary column for 2 minutes, and then washes the capillary column with a concentration of 100mg·ml -1 Carboxylated chitosan aqueous solution (weight-average molecular weight 200000) washes the capillary column for 10...

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Abstract

The invention relates to a protein adsorption resisting capillary column. A plurality of protein adsorption resisting covalent bonding coatings are modified on the inner wall of a capillary, and are formed in a manner that photosensitive diazo-resins and carboxylation chitosan are irradiated by ultraviolet rays to generate a light curing crosslinking reaction; and the thickness of each layer of the protein adsorption resisting covalent bonding coatings is 1.1-1.8nm. The preparation method of the protein adsorption resisting capillary column comprises the following steps of performing static electricity self-assembly on the photosensitive diazo-resins and the carboxylation chitosan, and performing light curing crosslinking, so as to obtain the protein adsorption resisting covalent bonding coatings. Through the adoption of the protein adsorption resisting capillary column disclosed by the invention, the separating efficiency of the protein adsorption resisting capillary column is notably improved, and the protein adsorption resisting property is also good; the preparing process of the method disclosed by the invention is simple and easy to operate, and the protein adsorption resisting capillary column is low in cost, good in repeatability, non-toxic, safe, environment-friendly, good in stability and good in protein adsorption resistance, and can be used for separation, detection and purification of substances including protein, enzymes, peptide chains, hormones and the like.

Description

technical field [0001] The invention relates to the technical field of capillary column separation, in particular to an anti-protein adsorption capillary column and a preparation method of the capillary column. Background technique [0002] In recent years, capillary electrophoresis technology has become more and more widely used in the analysis of proteins, biopolymers or drugs, and has become a modern and highly efficient means of separation. However, with the development of this technology, problems that need to be overcome There are also more and more problems, such as poor repeatability, significant peak tailing, short service life, adsorption of samples on the inner wall of the capillary, and so on. Some of the above problems can be effectively solved by changing the concentration and pH value of the buffer, or by modifying the capillary dynamic coating through non-covalent or covalent bonding. [0003] However, non-covalently bonded coatings generally suffer from sho...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447
CPCG01N27/44704
Inventor 于冰丛海林陈垚
Owner QINGDAO UNIV
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