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Temperature-sensitive protein molecular engram monolithic column and preparation method and application thereof

A technology of molecular imprinting and monolithic column, which is applied to the preparation method of peptides, chemical instruments and methods, and other chemical processes, can solve the problems of temperature-sensitive protein molecular imprinting monolithic columns, such as no relevant papers or patent reports, etc., to achieve Large-scale production, simple operation process, and the effect of improving the selectivity of removal

Inactive Publication Date: 2010-09-01
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there have been no relevant papers or patent reports at home and abroad on the preparation method and application of the temperature-sensitive protein molecularly imprinted monolithic column, which is an innovative research in theory and method

Method used

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  • Temperature-sensitive protein molecular engram monolithic column and preparation method and application thereof
  • Temperature-sensitive protein molecular engram monolithic column and preparation method and application thereof
  • Temperature-sensitive protein molecular engram monolithic column and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A western blot material, which can be prepared as follows:

[0032] (a) Add 1.2ml of silylating reagent to methanol and mix well. The volume ratio of silylating reagent to methanol is 3:1. Two silylating reagents, methyltrimethoxysilane and γ-methacryloxypropane The volume ratio of base trimethoxysilane is 7:1;

[0033] (b) Adding a catalyst with a concentration of 1.2mol / L to (a), and mixing evenly, wherein the ratio of the silylating agent to the catalyst is 4:1;

[0034] (c) pouring the liquid in (b) into the high performance liquid chromatography stainless steel column, sealing both ends of the mold, and placing it vertically at a temperature of 45° C. for 20 hours;

[0035] (d) After the reaction is over, the stainless steel column is connected to the high-performance liquid chromatography system, and the column body is washed with methanol to obtain a silica gel monolithic column skeleton; figure 1 It is a scanning electron micrograph of the cross-section of the...

Embodiment 2

[0044] A western blot material, which can be prepared as follows:

[0045] (a) Add 1.2ml of silylating reagent to ethanol and mix well. The volume ratio of silylating reagent to methanol is 2:1. Two silylating reagents, methyltriethoxysilane and γ-methacryloxy The volume ratio of propyltrimethoxysilane is 9:1;

[0046] (b) Add a catalyst with a concentration of 1.0mol / L to (a), and mix evenly, wherein the ratio of the silylating agent to the catalyst is 4:1;

[0047] (c) pouring the liquid in (b) into the capillary column, sealing both ends of the mold, and placing it vertically at a temperature of 47° C. for 30 hours;

[0048] (d) after the reaction is finished, the column body is washed with methanol to obtain a silica gel monolithic column skeleton;

[0049] (e) Dissolving the functional monomer and the cross-linking agent in the tris-hydrochloric acid (Tris-HCl) buffer solution of pH 7.10 of 10mM, the functional monomer in the buffer solution is NIPAAm (1.0290g), AAm ( ...

Embodiment 3

[0056] A western blot material, which can be prepared as follows:

[0057] (a) Add 1.2ml of silylating reagent to ethanol, mix well, the volume ratio of silylating reagent to methanol is 3:1, the volume of two silylating reagents tetraethoxysilane and aminopropyltrimethoxysilane The ratio is 5:1;

[0058] (b) adding a catalyst with a concentration of 0.8mol / L to (a), and mixing evenly, wherein the ratio of the silylating agent to the catalyst is 4:1;

[0059] (c) pouring the liquid in (b) into the capillary column, sealing both ends of the mold, and placing it vertically at a temperature of 50° C. for 12 hours;

[0060] (d) after the reaction is finished, the column body is washed with methanol to obtain a silica gel monolithic column skeleton;

[0061] (e) Dissolving the functional monomer and the cross-linking agent in the tris-hydrochloric acid (Tris-HCl) buffer solution of pH 7.10 of 10mM, the functional monomer in the buffer solution is NIPAAm (1.0513g), AAm ( 20.0mg) ...

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Abstract

The invention discloses a temperature-sensitive protein molecular engram monolithic column and a preparation method and application thereof, belonging to high molecular material preparation technology and application thereof in the field of protein separation and analysis, and particularly relating to a novel protein molecular engram monolithic column. A simple and moderate method is adopted to combine an inorganic silica gel monolithic column skeleton, protein serves as a template, and a temperature-sensitive functional monomer and cross-linking agent ingraft protein engram material on the surface of the inorganic silica gel so as to obtain the temperature-sensitive protein molecular engram monolithic column which has stronger template protein recognition capability. The temperature-sensitive protein molecular engram monolithic column prepared with the method of the invention combines the single identification characteristic of molecular engram with the characteristic of on-line separation of the monolithic column, has simple operation, moderate condition and low cost and provides a new method for removing high-abundance protein for the Journal of Proteome Research.

Description

【Technical field】 [0001] The invention belongs to polymer material preparation technology and its application in the field of protein separation and analysis, in particular to a novel protein molecularly imprinted monolithic column. 【Background technique】 [0002] Molecular imprinting technology (molecular imprinting technology) refers to the technology of preparing target molecules with specific recognition function, which is usually described as an artificial "lock" technology for manufacturing recognition "molecular keys". The basic principle is: provide suitable reaction conditions to make template molecules interact with functional monomers to form supramolecular complexes. The complexes form polymers under the action of cross-linking agents and initiators, and then remove template molecules under certain conditions. A molecularly imprinted polymer (molecularly imprinted polymer, MIP) with a recognition site complementary to the spatial structure of the template molecul...

Claims

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Application Information

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IPC IPC(8): B01J20/285B01J20/30C07K1/16
Inventor 张玉奎秦磊李文友何锡文
Owner NANKAI UNIV
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