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An affinity chromatography medium with hollow zinc oxide microspheres as a rigid matrix

A chromatographic medium, zinc oxide technology, applied in chemical instruments and methods, analytical materials, other chemical processes, etc., can solve the problems of high non-specific adsorption, poor biocompatibility, difficult affinity chromatographic medium, etc., to achieve large Effect of pressure tolerance range and strong mechanical strength

Active Publication Date: 2020-07-14
SUZHOU BOJIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the most widely used and commercialized matrices are non-rigid affinity chromatography media such as polydextrose, agarose, polycellulose, polyacrylic acid and polystyrene, which will deform to a certain extent under hydrostatic pressure. , will lead to increased pressure and clogging of the chromatographic column, which will result in a reduction or complete loss of the specificity and separation efficiency of the affinity chromatography medium, making it difficult to meet the large and fast operation requirements in the purification of biomacromolecules
However, rigid media such as silica, zirconia, alumina, and titania that can be used in other chromatography columns are difficult to be used as affinity chromatography media due to problems such as high non-specific adsorption and poor biocompatibility.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The preparation process is as follows:

[0023] (1) Place 20 g of polylactic acid-glycolic acid microspheres with a particle size of 50 to 100 μm in 100 mL of ethanol solution with a volume fraction of 50%, and magnetically stir at 35 to 40° C. for 30 minutes;

[0024] (2) Use 1mol / L sulfuric acid to adjust the pH value of the system to 2-3, add 20g of zinc sulfate powder, and magnetically stir at 30-35°C to dissolve the zinc sulfate powder;

[0025] (3) Add 2mol / L sodium hydroxide solution drop by drop while stirring magnetically at 30-35°C until the pH of the system is 9-9.5, continue magnetic stirring for 30 minutes and ultrasonication for 15 minutes, filter and rinse with deionized water, Dry to obtain polylactic acid-glycolic acid microspheres whose surface is covered with a zinc oxide shell;

[0026] (4) Place the microspheres obtained in step (3) in 20 mL of absolute ethanol at 20 to 25 ° C, add 5 mL of dioctyl sodium sulfosuccinate and 20 mL of 1,2-epoxybutane ...

Embodiment 2

[0031] The preparation process is as follows:

[0032] The protein A ligand in step (7) in Example 1 was replaced with the protein G ligand, and the rest of the preparation process was the same as in Example 1.

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PUM

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Abstract

The invention relates to an affinity chromatography medium using hollow zinc oxide microspheres as a rigid medium. An inner core of the affinity chromatography medium comprise the rigid medium and a non-rigid medium, the hollow zinc oxide microspheres serve as the rigid medium, and the non-rigid medium comprises a polylactic acid-glycolic acid copolymer, sodium dioctyl sulfosuccinate and 1,2-butylene oxide; the surface of the inner core is crosslinked with aminocellulose and is then activated by an epoxy group, and the epoxy group is connected to a ligand. The affinity chromatography medium not only has the advantages of relatively strong mechanical strength, relatively large pressure tolerance range and the like, but also has relatively good hydrophilicity and biocompatibility; meanwhile,the affinity chromatography medium is simple in production steps and low in production cost, and is conducive to large-scale popularization and application.

Description

technical field [0001] The invention relates to a chromatographic medium, in particular to an affinity chromatographic medium with hollow zinc oxide microspheres as a rigid matrix. Background technique [0002] The development of biotechnology has provided the possibility for the large-scale production of protein drugs with high added value. The industrialized large-scale purification of protein drugs, higher purity standards and economic cost requirements have promoted the great development of bioseparation technology. Liquid chromatography is an important unit in the biological separation process and plays an important role in the biopharmaceutical industry. One of the mechanisms of liquid chromatography is to adsorb various components in the sample through chemical or physical action. The specific action modes are: ion exchange chromatography, hydrophobic interaction chromatography, affinity chromatography, reversed phase chromatography and mixed mode layer analysis. A...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/26B01J20/30B01D15/38G01N33/558
CPCB01D15/3804B01J20/06B01J20/24B01J20/262B01J2220/4806B01J2220/4812B01J2220/4825G01N33/558
Inventor 瞿欢欢朱至放
Owner SUZHOU BOJIN BIOLOGICAL TECH