Immobilized metal chelate chromatography medium, and preparation method and application thereof

An immobilized metal, chelation chromatography technology, applied in chemical instruments and methods, separation methods, other chemical processes, etc., can solve the problems of increasing the preparation cost, cumbersome operation process, difficult quantitative control, etc. Good application prospects and the effect of increasing the quantity

Pending Publication Date: 2020-08-25
BEIJING INSTITUTE OF PETROCHEMICAL TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The operation process of this type of method is cumbersome, and an anhydrous environment is required in the process of introducing the initiator, the conditions are harsh, and it is difficult to quantitatively control each step, so it is necessary to add an excessive amount of reaction reagents, resulting in a certain amount of waste and increasing the preparation cost.

Method used

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  • Immobilized metal chelate chromatography medium, and preparation method and application thereof
  • Immobilized metal chelate chromatography medium, and preparation method and application thereof
  • Immobilized metal chelate chromatography medium, and preparation method and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0030] A preparation method for an immobilized metal chelate chromatography medium may comprise the following steps:

[0031] Step 1. Hydrolyzing the macroporous polymer microspheres under acidic conditions.

[0032] Step 2. Swelling the macroporous polymer microspheres treated in step 1 in the functional monomer solution.

[0033] Step 3. Stir and mix the initiator and the material treated in step 2 in a solvent, carry out graft polymerization reaction at a certain pH value and a certain temperature, and then wash away soluble substances with deionized water.

[0034] Step 4, putting the material treated in step 3 into a metal ion solution of a certain concentration and soaking for a certain period of time, thereby obtaining an immobilized metal chelation chromatography medium.

Embodiment approach

[0036] (1) In step 1, the macroporous polymer microspheres can be polyacrylate microspheres or polystyrene microspheres; the polyacrylate microspheres can be polyacrylate modified products, Derivative products or graft copolymers; the polystyrene microspheres can be polystyrene modified products, derivative products or graft copolymers; for example: the polyacrylate microspheres preferably adopt methacrylic acid shrinkage Copolymerized microspheres of glyceride and ethylene glycol dimethacrylate or self-polymerized microspheres of ethylene glycol dimethacrylate. This type of macroporous polymer microsphere can be prepared by atom transfer radical polymerization; this type of macroporous polymer microsphere is porous and interconnected, with a pore size range of 100-3000 nm and a particle size of 20-200 μm. The strength is above 1.0MPa, and the specific surface area is 5-200m 2 / g, using this kind of macroporous polymer microspheres with high mechanical strength, large pore si...

Embodiment 1

[0055] A kind of immobilized metal chelation chromatography medium, its preparation method can comprise the following steps:

[0056] Step A1, macroporous PGMA-EDMA microspheres are hydrolyzed under acidic conditions:

[0057] Accurately weigh 2.0 g of macroporous PGMA-EDMA microspheres into a 50 mL Erlenmeyer flask, then add 10 mL of dilute sulfuric acid solution with a concentration of 0.5 mol / L, and seal and vibrate for 3 h at room temperature in a vibrating shaker at 120 rpm. Suction filtration and washing with the same volume of deionized water, after washing, the solvent is suction filtered until the surface of the microspheres is neutral.

[0058] Step A2, preparation of composite monomer solution:

[0059] Accurately weigh sodium hydroxide and iminodiacetic acid (IDA), a tridentate chelating ligand, with a molar ratio of 2:1, and add an appropriate amount of water to completely dissolve it to obtain iminodiacetic acid sodium salt solution; slowly pour the iminodiacet...

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Abstract

The invention discloses an immobilized metal chelate chromatography medium, and a preparation method and application thereof. The preparation method comprises the following steps of: hydrolyzing macroporous polymer microspheres under an acidic condition; carrying out swelling treatment an obtained product in a functional monomer solution; stirring and mixing an obtained product and an initiator ina solvent; carrying out graft polymerization reaction at a certain pH value and a certain temperature; washing off soluble substances by using deionized water; and soaking an obtained product in a metal ion solution with a certain concentration for a certain period of time, so as to obtain the immobilized metal chelate chromatography medium. The preparation method is simple to operate and has nonon-harsh requirements for the environment; the number of functional groups on the surface of the material is effectively increased; the protein loading capacity of the medium is improved; and metal ion leakage is reduced. The prepared immobilized metal chelate chromatography medium is high in mechanical strength, low in non-specific adsorption, strong in specific binding force, high in protein binding loading capacity, high in operation flow rate and suitable for high-flux separation and purification of biomacromolecules.

Description

technical field [0001] The invention relates to the field of surface modification and graft modification of macroporous polymer chromatographic media, in particular to an immobilized metal chelating chromatographic media and its preparation method and application. Background technique [0002] With the rapid development of biopharmaceutical technology, there are more and more protein drugs, and the upstream and downstream processing of proteins are the key links in the manufacture of protein drugs. Although the upstream processing yield of protein has been greatly increased, the downstream processing and purification of protein is still the rate-limiting step, accounting for 50% to 90% of the total manufacturing cost of the product. Therefore, it is urgent to develop high-throughput, low-cost protein separation and purification methods. [0003] Immobilized metal chelation chromatography medium is a technique for the efficient separation of recombinant proteins. At present,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/286B01J20/30B01D15/38C08J9/40C08J9/36C08L33/14
CPCB01J20/286B01D15/38C08J9/405C08J9/36C08J2333/14B01D2015/3838
Inventor 张荣月侯恒磊靳海波何广湘
Owner BEIJING INSTITUTE OF PETROCHEMICAL TECHNOLOGY
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