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Preparation and use method of stereo floating protein molecule imprinting chromatographic stationary phase

A chromatographic stationary phase and molecular imprinting technology, which is applied in the field of chromatographic separation, can solve problems such as protein stereorecognition effects that are not considered, and achieve the effect of overcoming low adsorption capacity

Inactive Publication Date: 2012-11-28
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Traditional western imprinted polymers rely on imprinted sites to selectively recognize template proteins, without considering the three-dimensional recognition effect of proteins remaining in the polymer, so it is expected to remove the template to the greatest extent to obtain more imprinted site

Method used

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  • Preparation and use method of stereo floating protein molecule imprinting chromatographic stationary phase
  • Preparation and use method of stereo floating protein molecule imprinting chromatographic stationary phase
  • Preparation and use method of stereo floating protein molecule imprinting chromatographic stationary phase

Examples

Experimental program
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Embodiment 1

[0023] (1) Modification of the surface of silica gel: Weigh 1 g of non-porous silica gel (1.7 μm) into a 100 ml round bottom flask, add 40 mL of ultrapure water, cook in boiling water for 2 hours, cool, filter, rinse with methanol and dry overnight. Add 50mL of toluene (dry) to the dried silicon spheres, slowly add 2mL of 3-glycidoxypropyltrimethoxysilane dropwise, reflux at 120°C for 17h, cool and suction filter, wash with acetone and ethanol three times, and dry until use.

[0024] (2) Preparation of three-dimensionally floating hemoglobin-imprinted silicon spheres: Weigh 100 mg of the above materials into a round-bottomed flask, add 25 mL of 0.75 mol / L IDA solution, react in a water bath at 70°C for 17 hours, and wash with a large amount of water to remove the remaining IDA. Add 25mL 0.100mol / L CuSO4 aqueous solution, shake the reaction for 30min, and wash off the remaining CuSO4. Add 2 mL of hemoglobin (Hb, 5 mg / mL) in PBS solution (100 mM, pH=8.0), absorb for 10 min, and...

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Abstract

The invention belongs to the field of separating materials, and particularly relates to a preparation and use method of a protein molecule imprinting separating medium. A template protein molecule is connected to the surface of a matrix such as silica gel, a high molecule and the like through a physical or chemical method; a protein part combined with the surface of the matrix is further embedded by using a partial burying technology; unstable partially embedded proteins are cleared away; the residual proteins float on the surface of the matrix; and the residual proteins and imprinting points on the surface of the matrix achieve a stereo imprinting effect. Such a protein imprinting separating medium can perform high-selectivity separation on a template protein. The invention has the advantages of simple separating material preparation process and high selectivity.

Description

【Technical field】 [0001] The invention belongs to the field of chromatographic separation, in particular to a method for preparing and using a protein molecular imprinted separation medium. 【Background technique】 [0002] Molecular imprinting technology (MIT) refers to the technology of preparing polymers with selective recognition ability for a specific molecule. Template molecules interact with functional monomers to form supramolecular compounds, and polymers are formed under the action of cross-linking agents. When template molecules are removed under certain conditions, molecularly imprinted polymers (Molecular imprinting polymers, MIPs) can be obtained. [0003] The preparation methods of western imprinted polymers are generally divided into embedding method, surface imprinting method and epitope method. The surface imprinting method is an ideal method because its imprinting site is on or near the surface of the carrier particle, which has a faster association rate an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/286B01J20/30B01D15/08C07K1/16
Inventor 孟子辉雷雯张维冰薛敏
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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