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Protein-based magnetic agarose porous microspheres and preparation method thereof

A technology of agarose microspheres and agarose, applied in the field of magnetic agarose porous microspheres and its preparation, can solve the problems of long retention time of IgG and poor alkali resistance, achieve good anti-oxidation and alkali resistance, and improve alkali resistance , the effect of increasing the load

Pending Publication Date: 2022-06-03
苏州华诺生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] For this reason, the technical problem to be solved by the present invention is to overcome the problems of long IgG retention time and poor alkali resistance in the prior art

Method used

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  • Protein-based magnetic agarose porous microspheres and preparation method thereof
  • Protein-based magnetic agarose porous microspheres and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0028] A protein-based magnetic agarose porous microsphere and a preparation method thereof, specifically comprising the following steps:

[0029] 1. Pass 13.74g Fe 2 SO 4 ·7H 2 O, 24g FeCl 3 ·6H 2 O, 100 g water, room temperature N 2 Pass 20min, add 50gNH 3 ·H 2 O, stirred at room temperature for 2h to obtain nano Fe 3 O 4 .

[0030] 2. Take the above nano Fe 3 O 4 10 g, add 100 mL of ethanol, 2 mL of ammonia water, 2 g of TEOS, and stir at room temperature for 24 h to obtain silanol-modified Fe 3 O 4 .

[0031] 3. The above-mentioned silanol modified Fe 3 O 4 10 g was heated to 65 ℃, added 10 mL of 4% mass concentration agarose aqueous solution at 65 ℃, 30 g of paraffin oil at 65 ℃ and 1 g of Tween 20, and stirred at 500 rpm for 20 min. The temperature was lowered to 25°C to obtain magnetic agarose microspheres.

[0032] 4. Add 10 g of the above microspheres to 2 g of propylene glycol diglycidyl ester, add 800 μL of 48% NaOH, and react at 50° C. for 16 h to ...

Embodiment 2

[0036] A protein-based magnetic agarose porous microsphere and a preparation method thereof, specifically comprising the following steps:

[0037] 1. Pass 13.74g Fe 2 SO 4 ·7H 2 O, 24g FeCl 3 ·6H 2 O, 100 g water, room temperature N 2 Pass 20min, add 50gNH 3 ·H 2 O, stirred at room temperature for 2h to obtain nano Fe 3 O 4 .

[0038] 2. Take the above nano Fe 3 O 4 10 g, add 100 mL of ethanol, 2 mL of ammonia water, 2 g of TEOS, and stir at room temperature for 24 h to obtain silanol-modified Fe 3 O 4 .

[0039] 3. The above-mentioned silanol modified Fe 3 O 4 10 g was heated to 65 ℃, added 10 mL of 4% mass concentration agarose aqueous solution at 65 ℃, 30 g of cyclohexane at 65 ℃ and 1 g of Span 85, stirred at 500 rpm for 20 min, after the reaction was completed, at a rate of 5 ℃ / min The temperature was rapidly lowered to 25°C to obtain magnetic agarose microspheres.

[0040] 4. Add 10 g of the above microspheres to 2 g of propylene glycol diglycidyl ester,...

Embodiment 3

[0044] A protein-based magnetic agarose porous microsphere and a preparation method thereof, specifically comprising the following steps:

[0045] 1. Pass 13.74g Fe 2 SO 4 ·7H 2 O, 24g FeCl 3·6H 2 O, 100 g water, room temperature N 2 Pass 20min, add 50gNH 3 ·H 2 O, stirred at room temperature for 2h to obtain nano Fe 3 O 4 .

[0046] 2. Take the above nano Fe 3 O 4 10 g, add 100 mL of ethanol, 2 mL of ammonia water, 2 g of TEOS, and stir at room temperature for 24 h to obtain silanol-modified Fe 3 O 4 .

[0047] 3. The above-mentioned silanol modified Fe 3 O 4 10 g was heated to 65 ℃, added 10 mL of 4% mass concentration agarose aqueous solution at 65 ℃, 30 g of n-heptane at 65 ℃ and 1 g of Span 80, and stirred at 500 rpm for 20 min. After the reaction was completed, at a rate of 5 ℃ / min The temperature was rapidly lowered to 25°C to obtain magnetic agarose microspheres.

[0048] 4. Add 10 g of the above microspheres to 2 g of propylene glycol diglycidyl ester...

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Abstract

The invention relates to a protein-based magnetic agarose porous microsphere and a preparation method thereof, and belongs to the technical field of agarose microspheres. The preparation method comprises the following steps: firstly, uniformly mixing silicon hydroxyl modified magnetic nanoparticles, agarose, an oil phase material and an emulsifier, and carrying out stirring reaction at 60-70 DEG C for 15-25 minutes to obtain magnetic agarose microspheres; adding a cross-linking agent and an epoxy activator into the magnetic agarose microspheres under an alkaline condition, and reacting at 45-55 DEG C for 6-18 hours to obtain activated magnetic agarose microspheres; and finally, under an alkaline condition, adding protein A into the activated magnetic agarose microspheres, and reacting at 1-10 DEG C for 20-28 hours to obtain the protein-based magnetic agarose microspheres. The magnetic agarose porous microspheres have alkali resistance, the alkali resistance range pH is 10-14, the agarose microspheres can keep chemical stability, and the maximum immunoglobulin IgG binding capacity can reach 60 mg / mL.

Description

technical field [0001] The invention relates to the technical field of agarose microspheres, in particular to a protein-based magnetic agarose porous microsphere and a preparation method thereof. Background technique [0002] With the development of biotechnology, new antibodies continue to emerge. Generally speaking, the development process of new antibodies needs to go through a high-throughput screening process. In this process, purification and cleaning are essential, and alkaline solvents such as NaOH are commonly used. detergent. Agarose microspheres have good biocompatibility and become the stationary phase matrix in the most widely used separation column in the field of biological protein separation. However, the alkali resistance of the currently used magnetic agarose screening column is limited, and high-throughput screening cannot be achieved with automated equipment for sample processing. After several times of washing with NaOH during the purification process, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J13/14
CPCB01J13/14
Inventor 不公告发明人
Owner 苏州华诺生物科技有限公司
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