Preparation method of magnetic beads for protein purification

A protein purification and magnetic bead technology, applied in the biological field, can solve problems such as protein denaturation, and achieve rapid separation, convenient separation, and easy operation

Active Publication Date: 2017-01-04
RUBYBERRIES BIOTECHNOLOGIES CO LTD
View PDF6 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This treatment method is more effective for the recovery and adsorption of heavy metal ions in industrial wastewater pollution,

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Preparation method of magnetic beads for protein purification
  • Preparation method of magnetic beads for protein purification

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0021] Example 1

[0022] (1) Take 9g of chitosan, add 900ml of pure water, add 3ml of glacial acetic acid under stirring, fully stir to dissolve, cool to 4°C to prepare a chitosan solution; (2) Add 750g of magnetic powder to a 10L reaction system , 1875ml of distilled water, add 750ml of chitosan solution prepared in step (1), add 37.5ml of cold 4% glyoxal solution under high-speed stirring, and mix well; (3) add 0.2mol / L K 2 HPO 4 220ml solution, stirring at high speed for 2h; (4) adding (NH 4 ) 2 SO 4 40g, while dynamically adjusting and maintaining the pH of the solution at about 8.5 with 1mol / L NaOH solution, stirring for 2h; (5) adding NaBH 4 10g, stirred for 2h, remove the supernatant, wash twice with 5L distilled water; (6) add 66g of sodium chloroacetate, stir for 2h at 85°C and pH 8.5; (7) the mixture obtained by the above steps, Wash with distilled water twice to obtain magnetic beads for protein purification.

[0023] Take 0.3g (wet) of the above-mentioned magnetic b...

Example Embodiment

[0024] Example 2mol / L

[0025] (1) Take 9g of chitosan, add 900ml of pure water, add 3ml of glacial acetic acid under stirring, fully stir to dissolve, cool to 4℃ to prepare a chitosan solution; (2) Add 750g magnetic powder to a 10L reaction system , 1875ml of distilled water, add 750ml of chitosan solution prepared in step one, add 37.5ml of cold 4% glutaraldehyde solution under high-speed stirring, and mix well; (3) add 0.2m / L of K 2 HPO 4 220ml solution, stirring at high speed for 2h; (4) adding (NH 4 ) 2 SO 4 40g, while dynamically adjusting and maintaining the pH of the solution at about 8.5 with 1mol / L NaOH solution, stirring for 2h; (5) adding NaBH 4 10g, stirred for 2h, remove the supernatant, wash twice with 5L distilled water; (6) Add 66g of sodium chloroacetate at 85°C and pH 8.5, stir for 2h; (7) The mixture obtained by the above steps, Wash with distilled water twice to obtain magnetic beads for protein purification.

[0026] The protein purification effect was teste...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a preparation method of magnetic beads for protein purification. Chitosan and magnetic powder are used as raw materials, core-shell structure particles of crosslinked chitosan coated magnetic powder are formed under the effect of a cross-linking agent, and the magnetic beads for protein purification are formed under reduction and carboxymethylation effects. The magnetic beads for protein purification are used for separation and purification of protein with his-tag, and the separation efficiency is greatly improved. The problem of protein denaturation possibly occurs during elution is avoided, and the protein separating efficiency can be improved. In addition, the preparation method is convenient and quick to operate, is suitable for separation of a turbid protein solution and is especially suitable for great industrial production.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing magnetic beads used in protein separation and purification. Background technique [0002] The separation and purification of proteins mainly include methods such as salting out, centrifugation, gel electrophoresis and affinity chromatography. Among them, salting out, centrifugation and gel electrophoresis are limited by factors such as high cost and low efficiency, and their applications are relatively limited. Affinity chromatography is to make an affinity molecule with a special structure into a solid-phase adsorbent and place it in a chromatography column. When the protein mixture to be separated passes through the chromatography column, the protein with affinity with the adsorbent will be It will be adsorbed and retained in the chromatographic column, and can be separated and purified after being eluted. At present, for the separation and pu...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): B01J13/14C07K1/14
CPCB01J13/14C07K1/14
Inventor 金彩科郑诚
Owner RUBYBERRIES BIOTECHNOLOGIES CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap