Composite filler and preparation method thereof

A composite filler and composite matrix technology, which is applied in the preparation methods of peptides, chemical instruments and methods, ion exchange, etc., can solve the problems of affecting the loading of the filler and the flow rate, slow flow rate affecting purification efficiency, and low protein purification efficiency. Improve purification efficiency and effect, stable physical and chemical properties, and improve separation efficiency

Inactive Publication Date: 2018-11-16
浙江月旭材料科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the domestic packing capacity is low, generally 30mg/ml, the slow flow rate affects the purification efficiency, and the protein recovery rate is low, and the protein activity is low
[0012] At present, domestic manufacturers can also produce various types of purification fillers, but there are still certain limitations. For example, there are thousands of types of proteins, and some proteins also have ion exchange properties and hydrophobic pro...

Method used

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  • Composite filler and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Preparation of composite filler:

[0054] (1) Preparation of composite base balls: Take 100g of cellulose, add it to a 1L three-necked flask, add 200ml of deionized water, put the three-necked flask into a constant temperature water bath, and stir at 40°C and 200rpm for 0.5h; The temperature of the pot is raised to 92°C, add 17g of agarose to the three-necked flask, and stir for 2 hours; continue to add 1g of disodium hydrogen phosphate and 1g of sodium sulfate, so that the pH of the mixture is 9-10; take the agarose and cellulose mixture Measure the viscosity with a viscometer. When the viscosity is 600-800 Pa.s, stop heating and cool down the agarose-cellulose mixture to 50°C. Take 100ml of toluene and put it into another 500ml three-necked flask, put the three-necked flask into a water bath, stir at 50°C and 100rpm for 0.5h, add 1ml of Span 80, and continue stirring for 1h. Slowly pour the cooled agarose cellulose mixture into the toluene Span 80 mixture, and contin...

Embodiment 2

[0060] Preparation of composite filler:

[0061] (1) Preparation of composite base balls: Take 100g of cellulose, add it to a 1L three-necked flask, add 300ml of deionized water, put the three-necked flask into a constant temperature water bath, and stir at 50°C and 300rpm for 1 hour; put the water bath The temperature of the temperature rises to 98°C, add 20g of agarose to the three-necked flask, and stir for 3 hours; continue to add 1.5g of disodium hydrogen phosphate and 1.5g of sodium sulfate, so that the pH of the mixture is 9-10; take the agarose cellulose and mix Measure the viscosity of the liquid with a viscometer. When the viscosity is 600-800 Pa.s, stop heating and cool down the agarose-cellulose mixture to 60°C. Take 300ml of toluene and put it into another 500ml three-necked flask, put the three-necked flask into a water bath, stir at 60°C and 150rpm for 1h, add 3ml of Span 80, and continue stirring for 2h. Slowly pour the cooled agarose cellulose mixture into th...

Embodiment 3

[0067] Preparation of composite filler:

[0068] (1) Preparation of composite base balls: Take 100g of cellulose, add it to a 1L three-necked flask, add 400ml of deionized water, put the three-necked flask into a constant temperature water bath, and stir at 60°C and 200rpm for 2 hours; put the water bath The temperature of the temperature rises to 100°C, add 25g of agarose to the three-necked flask, and stir for 4 hours; continue to add 1.5g of disodium hydrogen phosphate and 1.5g of sodium sulfate, so that the pH of the mixture is 9 to 10; take the agarose cellulose and mix Measure the viscosity of the liquid with a viscometer. When the viscosity is 600-800 Pa.s, stop heating and cool down the agarose-cellulose mixture to 70°C. Put 600ml of toluene into another 1L three-necked flask, put the three-necked flask into a water bath, stir at 60°C and 150rpm for 1h, add 3ml of Span 80, and continue stirring for 3h. Slowly pour the cooled agarose cellulose mixture into the toluene ...

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Abstract

The invention discloses composite filler. The composite filler contains a compound substrate mainly prepared from agarose and cellulose and composite ligands crosslinked with the composite substrate,wherein the composite ligands comprise ion exchange ligands and affinity ligands, the ion exchange ligands are sodium metabisulfite and aminobenzene, and the affinity ligands are hydroxyapatite, protein A or recombinant protein A. The composite filler integrates characters of multiple substrates and multiple ligands, has the characteristic of high protein loading capacity, and can increase workingflow velocity, thereby effectively improving purification efficiency and effect of protein.

Description

【Technical field】 [0001] The invention belongs to the technical field of protein purification packing, and in particular relates to a composite packing and a preparation method thereof. 【Background technique】 [0002] Foreign filler production technology and filler diversity research and development have developed to a very high level. In 1959, the team led by the Nobel laureate Professor Arne Tiselius formally proposed the concept of gel filtration filler, which opened the door to a new method of protein purification. To date, dozens of different types of fillers have been developed. The filler composition is mainly divided into two parts: the first part is the matrix, which is spherical particles cross-linked by polysaccharides or polymers. The size of the base ball is about 90um, and there are a large number of small holes on the surface and inside for the protein to enter the inside. At the same time, the surface and interior of the base ball are rich in hydroxyl group...

Claims

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Application Information

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IPC IPC(8): B01D15/08C07K1/16
CPCB01D15/08C07K1/165
Inventor 赵培
Owner 浙江月旭材料科技有限公司
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