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A functional composite metal oxide material and its preparation method and application

A composite metal and oxide technology, applied in the field of cobalt, nickel, manganese composite metal oxide materials, can solve the problem of high cost, achieve the effect of improving the utilization rate, the method is simple and easy, and the mass production is convenient

Inactive Publication Date: 2018-08-28
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Literature (L. Zhang, X. Zhu, D. Jiao, Y. Sun and H. Sun. Efficient purification of His-tagged protein by superparamagnetic Fe 3 o 4 / Au–ANTA–Co 2+ nanoparticles.Mat.Sci.Eng.C-Mater.,2013,33,1989-1992) proposed Co 2+ The ability to bind histidine is better than that of Ni 2+ , but the cost is higher

Method used

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  • A functional composite metal oxide material and its preparation method and application
  • A functional composite metal oxide material and its preparation method and application
  • A functional composite metal oxide material and its preparation method and application

Examples

Experimental program
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Embodiment 1

[0032] Nickel, cobalt, manganese composite metal oxide material MnCoNiO 4 The synthesis of comprises three steps of co-precipitation, grinding and high temperature treatment, and the specific preparation method is as follows:

[0033] (1) Add 10mmol NiSO to a 1000mL beaker 4 ·6H 2 O. CoSO 4 ·7H 2 O and MnSO 4 ·H 2 O, then add 500 mL of deionized water until completely dissolved. An appropriate amount of ammonia water was added to adjust the pH of the solution to 10. Prepare 200 mL of a sodium carbonate solution with a concentration of 27.5 μg / mL, and then add it dropwise to the metal salt solution under heating and stirring at 70°C until the precipitation is complete. Wash the precipitate 3 times with deionized water, filter and dry the precipitate in a drying oven at 70°C for 10 hours;

[0034] (2) Use an agate mortar to fully grind the precipitate obtained in step (1) for 1 hour;

[0035] (3) Put the fully ground precipitate obtained in step (2) into a crucible, then ...

Embodiment 2

[0038] Nickel, cobalt, manganese composite metal oxide material MnCoNiO 4 The specific steps for isolating the hexahistidine-labeled yellow fluorescent protein in the enrichment solution are as follows:

[0039] 200 μg MnCoNiO 4 The microspheres were mixed with 500 μL of 20 μg / mL hexahistidine-labeled yellow fluorescent protein solution at room temperature for 5 minutes, centrifuged at 10,000 rpm for 2 minutes, and the supernatant was discarded. Then, 200 μL of phosphate buffer solution (pH=7.40) was added to the centrifuge tube, mixed for 1 min, and the supernatant was discarded after centrifugation. This washing process was repeated 2 times. Finally, 20 μL of 1 mol / L imidazole solution was added to the bottom of the centrifuge tube, rotated and mixed for 5 minutes, centrifuged at 10,000 rpm for 2 minutes, and the supernatant was retained. The supernatant is the composite metal oxide material MnCoNiO 4 The resulting hexahistidine-tagged yellow fluorescent protein solution...

Embodiment 3

[0042] Nickel, cobalt, manganese composite metal oxide material MnCoNiO 4 Separation and purification of the hexahistidine-labeled green fluorescent protein in the cell lysate, the specific steps are as follows:

[0043] The Escherichia coli expressing the hexahistidine-tagged green fluorescent protein was ultrasonically lysed, then centrifuged at a centrifugation rate of 12000 rpm for 20 min, and the supernatant was retained. Then 200 μg MnCoNiO was added to 200 μL supernatant 4 The microspheres were rotated and mixed for 5 minutes at room temperature, centrifuged at 10000 rpm for 2 minutes, and the supernatant was discarded. Then add 200 μL of 10 mmol / L imidazole solution to the centrifuge tube, mix for 1 min, and discard the supernatant after centrifugation. This washing process was repeated 3 times. Finally, 20 μL of 1 mol / L imidazole solution was added to the bottom of the centrifuge tube, rotated and mixed for 5 minutes, centrifuged at 10,000 rpm for 2 minutes, and th...

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Abstract

The invention discloses a functional composite metal oxide material, a preparation method and application thereof. According to the invention, coprecipitation method and high temperature solid state method are combined organically, and the precipitation rate and reaction temperature are reasonably controlled to prepare size controllable nickel, cobalt and manganese composite metal oxide nano-micron scale microspheres with regular shape. Based on the chelation of metal ions and histidine, histidine-tagged fusion protein can be adsorbed on the surfaces of the microspheres, and then can be eluted quickly with eluent. The preparation method of the microsphere material is simple and feasible, low in cost, and convenient for batch production. The prepared nickel, cobalt and manganese composite metal oxide nano-micron scale microsphere material has uniform and controllable shape, can be used for fast and efficient separation and purification of histidine-tagged fusion protein in complex biological samples, and can realize repeated cycle use, thus having very high commercial application prospect.

Description

technical field [0001] The invention relates to a functional composite metal oxide material, in particular to a nickel, cobalt and manganese composite metal oxide material, a preparation method of the material and its application in the separation and purification of hexahistidine fusion protein. Background technique [0002] Fusion tag technology emerged at the end of the 20th century and is a recombinant DNA technology based on a reporter gene. The rapid development of this technology makes the purification of recombinant proteins more efficient and convenient, and it is widely used in many fields such as life sciences and chemistry. Currently commonly used protein purification tags include hexahistidine, glutathione sulfhydryl transferase, and epitope tags, among which hexahistidine fusion proteins are the most commonly used. A large number of fusion proteins are expressed by Escherichia coli, etc., and then the expressed hexahistidine fusion protein is separated and pur...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/06B01J20/30C07K19/00C07K1/22
Inventor 白玉祁晓月刘虎威
Owner PEKING UNIV
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