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Method for packing histidine-tagged protein with nano silica

A nano-silica, histidine-labeled technology, applied in the preparation methods of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problems of changing the chargeability of proteins, low encapsulation efficiency, and easy leakage of nano-silica. , to achieve good thermal stability, not easy to enzymatic hydrolysis, good stability

Inactive Publication Date: 2012-07-04
SHANGHAI UNIV
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Problems solved by technology

The nano-silica wrapped by the traditional method is easy to leak in the solution, and the wrapping efficiency is low. This method takes advantage of the characteristics of the histidine-tagged protein and selects a suitable metal ion that can be complexed with the imidazole group to complex with the histidine-tagged protein. , changed the charge of the protein, and prepared stable, monodisperse nano-silica particles

Method used

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  • Method for packing histidine-tagged protein with nano silica
  • Method for packing histidine-tagged protein with nano silica
  • Method for packing histidine-tagged protein with nano silica

Examples

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Embodiment

[0021] Take histidine-tagged green fluorescent protein as an example. Mix 7.5 ml of cyclohexane, 1.77 ml of polyethylene glycol octylphenyl ether, and 1.8 ml of n-hexanol, add 300 μl of a mixture of calcium chloride and histidine-tagged green fluorescent protein, and then add 100 Microliter tetraethyl orthosilicate, stir evenly with magnetic force, and finally use 60 microliters of ammonia water to initiate polymerization. After stirring and reacting at room temperature for 24 hours, add 20 milliliters of acetone to precipitate the nanospheres, wash several times to remove impurities such as surfactants, and you are done. Nano-silica particles containing histidine-tagged green fluorescent protein were obtained.

[0022] The transmission electron microscope (TEM) diagram of the nano-silica particles comprising hischloroic acid-labeled green fluorescent protein obtained in this embodiment is shown in figure 1 .

[0023] from figure 1 It can be seen that the nano-silica partic...

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Abstract

The invention relates to a method for packing histidine-tagged protein with nano silica, belonging to the technical field of nano inorganic material and biomacromolecule chemistry. The method comprises the following key points: mixing cyclohexane, polyethylene glycol phehexyl alcoholctane and hexyl alcohol uniformly by a certain proportion to obtain a microemulsion mixed liquor system, adding mixture of metal ion water-soluble salt and histidine-tagged protein, then adding a certain amount of tetraethoxysilane, stirring uniformly under magnetic force, adding a small amount of ammonia water totrigger polymerization, stirring at room temperature for full reaction, adding acetone as a demulsifier to enable the nanospheres to precipitate, and cleaning repeatedly to remove surfactants and other impurities to obtain nano silica particles containing histidine-tagged protein. The nano silica particles containing histidine-tagged green fluorescent protein prepared by the method have good heatstability and other sound performances, and can not be decomposed by enzyme easily; and the fluorescent light can not be quenched easily.

Description

technical field [0001] The invention relates to a method for wrapping a histidine-tagged protein with nano silicon dioxide, which belongs to the technical field of nano inorganic materials and biopolymer chemistry. Background technique [0002] Histidine-tagged protein is a commonly used protein in protein purification. The purification efficiency of protein tagged with histidine is greatly increased, and the protein characteristics are basically unchanged. The application of nano-silica to coat proteins is a cross-research direction combining nanotechnology and biotechnology. The nano-silica wrapped by the traditional method is easy to leak in the solution, and the wrapping efficiency is low. This method takes advantage of the characteristics of the histidine-tagged protein and selects a suitable metal ion that can be complexed with the imidazole group to complex with the histidine-tagged protein. , changed the charge of the protein, and prepared stable, monodisperse nano-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14G01N21/64G01N33/68
Inventor 曹傲能叶张梅蔡正伟董二亚刘刚波王海芳
Owner SHANGHAI UNIV
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