Histidine tag protein affinity purification material and application thereof

A histidine tag and protein technology, which is applied in the preparation of functional biological materials and in the field of protein recognition and purification, can solve the problems of lower purification purity of histidine tag, increase of purification cost, and low purification recovery rate, and achieve reduction The interference of strong non-specific adsorption, the effect of reducing the leakage of metal ions and improving the purification purity

Inactive Publication Date: 2017-03-15
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The adsorption of these proteins will lead to a significant reduction in the purity of histidine tag purification, and secondary or even tertiary purification must be carried out in order to achieve the required purity (J.K.Shea, etal.J.Am.Chem.Soc., 2014,136,1194- 1197), the more purification steps, the lower the purification recovery rate, and the corresponding purification cost is also greatly increased

Method used

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  • Histidine tag protein affinity purification material and application thereof
  • Histidine tag protein affinity purification material and application thereof
  • Histidine tag protein affinity purification material and application thereof

Examples

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Effect test

Embodiment 1

[0037] Based on IDA and Ni 2+ Preparation of histidine-tagged molecularly imprinted silica material (MIP1) for chelation

[0038] use Monodisperse silica nanoparticles were prepared by method, and surface-modified IDA and Ni were prepared by reference method 2+ The matrix material (L.Zhang, et al.Chem.Commun., 2011,47,3969-3971; L.Zhang, et al.Anal.Chem., DOI:10.1021 / ac5047246): 5g silica nanoparticles were added In 100mL GLYMO-IDA solution, react at 65°C for 24 hours to modify the IDA group on the surface of the silica material. 5 g of the above materials were added to 150 mL of methanol, followed by 10 mL of γ-MAPS, and refluxed at 90° C. for 15 hours to modify γ-MAPS on the surface of the matrix material so as to be imprinted as a matrix material. Then 100 mg of the above material was redispersed in 1 mL of 12 mg / mL template molecule histidine tag (HHHHHH) aqueous solution (the mass ratio of template molecule to the above matrix material was 120:1000), and incubated at ...

Embodiment 2

[0040] The non-imprinted material 1 (NIP1) was prepared by the same method as in Example 1, but without adding a template molecule histidine tag (HHHHHH).

Embodiment 3

[0042] Using the same method as in Example 1, only metal ions were immobilized on the matrix material without subsequent imprinting process to prepare non-imprinted material 2 (NIP2).

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Abstract

The invention relates to a histidine tag protein affinity purification material. The histidine tag protein affinity purification material is prepared from a molecular imprinting material based on a metal chelation function, an immobilized metal chelation affinity chromatography material (IMAC) utilized as a matrix material and a histidine tag utilized as a template module by using a molecular imprinting technique. The histidine tag protein affinity purification material combines the good affinity of the metal chelation function and the high selectivity of the molecular imprinting material, and can be further applied to high-purity purification of the histidine tag protein.

Description

technical field [0001] The invention belongs to the preparation of functional biological materials and their application in protein recognition and purification, specifically to the preparation of a new type of molecularly imprinted material based on metal chelation and its use as an affinity purification material for histidine tags Protein identification and purification. Background technique [0002] Recombinant protein technology is a very important technology that can obtain a large number of target proteins, and plays a very important role in protein crystallization, protein drug, protein interaction and structural proteomics research (T.Hyeon, et al.Adv. Mater., 2010, 22, 57–60; A.S. Robinson, et al. Biotechnol. J., 2012, 7, 620–634). From the basic research of protein structure and function to the development of functional protein expression and purification process, affinity tag has become an important and effective tool for recombinant protein purification, in whic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/286B01J20/30B01D15/08C07K1/22
Inventor 张丽华李森武杨开广赵宝锋李潇闵一张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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