Magnetic microsphere as well as preparation and preparation method thereof used for purifying histidine tag protein

A technology of histidine labeling and magnetic microspheres, applied in the field of magnetic microspheres, can solve the problems of cumbersome application procedures, difficult fast adsorption and movement, low response, etc., achieve efficient chelation of metal ions, and simple purification and separation process , Guarantee the effect of purification efficiency

Active Publication Date: 2010-06-23
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology uses tiny particles called magnetite microparticles (MMP), instead of conventional chemical reagents like EDTA, DTTN, and others that are commonly found in biological samples such as blood plasma. These MMP have been shown to quickly separate certain types of proteins from their resting state without losing them when exposed to strong electromagnetic fields. They also allow for easy removal of excessive charged residual charges during processing steps. Overall this technical effect provides improved methods for removing unwanted substances while maintaining important functions of these materials.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the performance and stability of enzymatic catalysis systems used in separations between different types of substances like peptides, nucleic acids, metabolites, hydroxyproline, urocanamines, crustones, ferrules, silver nitrate loaded onto silica gel surfaces, and insolvent precipitation techniques. These improvements require efficient methods for preparing and controlling the properties of certain compounds attached at the binding sites within porous supports called media columns.

Method used

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  • Magnetic microsphere as well as preparation and preparation method thereof used for purifying histidine tag protein
  • Magnetic microsphere as well as preparation and preparation method thereof used for purifying histidine tag protein
  • Magnetic microsphere as well as preparation and preparation method thereof used for purifying histidine tag protein

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Embodiment 1

[0037] (1) The method proposed in the Chinese invention patent ZL 2006100279617 was used to prepare magnetic silica microspheres with a particle size of 200nm in the surface-coated silica shell. Freeze-dry the silica microspheres, weigh 6g of freeze-dried magnetic particles, add them to a 100mL three-necked flask, and add 60mL of toluene to the flask; start stirring, put them in an oil bath at 120°C, and add them after reflux for 30 minutes 5 g of γ-aminopropyltrimethoxysilane was refluxed for 2 h. After the reaction was finished, ultrasonicated for 30 seconds, poured into a beaker, and separated under the assistance of a strong magnetic field, then added 50mL of toluene, ultrasonicated for 1 minute, and separated and washed by a strong magnetic field. After repeated operations three times, the obtained amino-modified magnetic microspheres Put it into a vacuum drying oven at 120°C for a vacuum reaction for 12 hours, and finally place the obtained magnetic microspheres in a sam...

Embodiment 2

[0044] (1) The method proposed in Chinese invention patent ZL 2006100279617 was used to prepare magnetic silica microspheres coated with a silica shell. The difference is that the inner core of the magnetic particle aggregates in the microspheres is 900nm, and the silicon oxide shell coating method is the same as that of the Chinese invention patent ZL 2006100279617, so the particle size of the finally obtained silicon oxide magnetic microspheres is 1000nm. Freeze-dry the silica microspheres, weigh 6g of freeze-dried magnetic particles, add them to a 100mL three-necked flask, and add 20mL of toluene to the flask; stir, put them in an oil bath at 120°C, reflux for 30 minutes, and then add γ - Aminopropyltriethoxysilane 1g, reflux reaction for 2h. After the reaction was finished, ultrasonicated for 30 seconds, poured into a beaker, and separated under the assistance of a strong magnetic field, then added 50mL of toluene, ultrasonicated for 1 minute, and separated and washed by a...

Embodiment 3

[0051] (1) The method proposed in Chinese invention patent ZL 2006100279617 was used to prepare magnetic silica microspheres coated with a silica shell. The difference is that the inner core of the magnetic particle aggregates in the microspheres is 900nm, and the silicon oxide shell coating method is the same as that of the Chinese invention patent ZL 2006100279617, so the particle size of the finally obtained silicon oxide magnetic microspheres is 1000nm. Freeze-dry the silicon oxide microspheres, weigh 6g of freeze-dried magnetic particles, add them to a 100mL three-necked flask, and add 20mL of toluene to the flask; stir, put them in an oil bath at 120°C, reflux for 30 minutes, and then add N -0.5 g of β(aminoethyl)-γ-aminopropyltrimethoxysilane, reflux for 2 hours. After the reaction was finished, ultrasonicated for 30 seconds, poured into a beaker, and separated under the assistance of a strong magnetic field, then added 50mL of toluene, ultrasonicated for 1 minute, and ...

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Abstract

The invention discloses a magnetic microsphere as well as preparation and preparation method thereof used for purifying histidine tag protein in the technical field of biological materials; wherein the magnetic microsphere includes 50wt percent to 80wt percent of ferroferric oxide nanoparticles, 40wt percent to 15wt percent of monox shell as well as 5wt percent to 10wt percent of hyperbranched polymer molecule with a poly carboxyl structure. The molecular structure of the hyperbranched polymer molecule with a poly carboxyl structure is as follows: wherein R refers to - (CH2CH2O-) 3-CH2CH2-. The invention adopts the magnetic microsphere with a quick magnetic response as a solid support, adopts the hyperbranched polymer molecule with a poly carboxyl structure to decorate on the surface of the magnetic microsphere to obtain a magnetic carrier material which can be effectively chelated with transition metal ions, and finally realizes the general goal of quickly, effectively and simply purifying and separating histidine tag protein.

Description

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Claims

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Application Information

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Owner SHANGHAI JIAO TONG UNIV
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