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MA/AA/MMA copolymer resin for biological macromolecule concentration and its preparation method

A biomacromolecule and copolymer resin technology, applied in the field of copolymer resin, can solve the problems of large sample loss, difficult cleaning, easy rupture of dialysis membrane, etc., and achieve the effect of simple manufacture and use, difficult cleaning and high price

Inactive Publication Date: 2005-06-08
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to propose a kind of acrylamide / acrylic acid / methyl methacrylate copolymer resin (MA / AA / MMA copolymer resin) and its application method for the concentration of biological macromolecules, and apply superabsorbent resin technology to biological macromolecules Concentration and dehydration to overcome the fact that the existing precipitation and adsorption methods of biomacromolecule concentration are easy to affect the properties of the sample. The buffer solution of the dialysis method needs to be replaced many times and the dialysis membrane is easily broken. Ultrafiltration is not suitable for the case of a small amount of sample. Moreover, some proteins may undergo irreversible denaturation, ultrafiltration membranes are expensive and difficult to clean, and the sample loss is large. The vacuum distillation method is easy to denature the protein by heat, and the freeze-drying method is difficult to operate and will cause a decrease in protein activity.

Method used

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  • MA/AA/MMA copolymer resin for biological macromolecule concentration and its preparation method
  • MA/AA/MMA copolymer resin for biological macromolecule concentration and its preparation method
  • MA/AA/MMA copolymer resin for biological macromolecule concentration and its preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0014] Embodiment 1: prepare MA / AA / MMA copolymer resin and be used for concentrating bovine serum albumin

[0015] First, use a balance to weigh 90g of AA liquid, 10g of MA solid, and 30g of sodium hydroxide solid; dissolve with 500mL of distilled water. Use a 50mL pipette to pipette 25mL of the above solution. Use a 1mL pipette to add 1mL of MMA liquid, and oscillate to suspend (ultrasonic oscillation method can be used here). The poly AA homopolymer superabsorbent resin has high water absorption and poor salt resistance, while the polyMA homopolymer super absorbent resin has low water absorption but strong salt tolerance, and most biological macromolecules are dissolved in salt solution , so it can be cross-linked with MA / AA monomers. Since both MA and AA monomers are hydrophilic compounds, they will absorb biomacromolecules and cause unnecessary losses, so the hydrophobic compound MMA is added according to the maximum suspension amount for copolymerization to obtain a new...

Embodiment 2

[0016] Embodiment 2: prepare another kind of MA / AA / MMA copolymer resin concentrated bovine serum albumin BSA concentration experiment

[0017] First, use a balance to weigh 90g of AA liquid, 20g of MA solid, and 45g of sodium hydroxide solid; dissolve with 300mL of distilled water. Use a 20mL pipette to pipette 15mL of the above solution. Add 1mL of MMA liquid with a 1mL pipette and shake to suspend. After mixing, use a syringe to draw the mixed solution into a medical drip tube with a diameter of 3 mm, and seal both ends of the plastic tube with iron clips. Immediately place the dropper filled with the mixed monomers under Co60γ-ray irradiation at 125Gy / min for 50min. Take 10 mL of the BSA solution to be concentrated, pipette 20 μL into EP tube 1, and mark it as “1×”. Cut the 4cm-long resin, peel off the outer tube wall, put it into the solution to be concentrated (15mL centrifuge tube), and let it stand at 4°C. When the remaining solution volume is observed to be 5 mL, t...

Embodiment 3

[0018] Embodiment 3: adopt copolymer resin of the present invention to concentrate bovine serum albumin experiment

[0019]First, weigh 90g AA liquid, 8g MA solid, and 40g sodium hydroxide solid with a balance; dissolve with 400mL distilled water. Use a 20mL pipette to pipette 20mL of the above solution. Use a 1mL pipette to add 1mL of MMA liquid, and oscillate to suspend (ultrasonic oscillation method can be used here). After mixing, use a syringe to draw the mixed solution into a medical drip tube with a diameter of 3 mm, and seal both ends of the plastic tube with iron clips. Immediately place the dropper filled with the mixed monomers under Co60γ-ray irradiation at 125Gy / min for 50min. Take 10 mL of the concentrated BSA solution, pipette 20 μL into EP tube 1, and mark it as “1×”. Cut the 4cm-long resin, peel off the outer tube wall, put it into the solution to be concentrated (15mL centrifuge tube), and let it stand at 4°C. When the remaining solution volume is observe...

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Abstract

A process for preparing the MA / AA / MMA copolymer resin used for concentrating the large biological molecular material includes such steps as proportionally mixing AA liquid, MA solid sodium hydroxide and distilled water, proportionally adding MMA, waving, and three element polymerizing under the triggering of gamma ray. Its advantages are no influence to specimen and low cost.

Description

Technical field: [0001] The invention belongs to the technical field of copolymer resins for absorbing and concentrating biological macromolecules, in particular to radiation synthesis of MA / AA / MMA copolymer resins and methods. Background technique: [0002] During the preparation of biomacromolecules, the concentration of the sample becomes very low due to column purification, making it difficult to conduct biochemical experiments directly. For the purpose of preservation and identification, concentration and dehydration are often required. [0003] According to "Principles and Applications of Biochemical Technology" edited by Zhao Yongfang (Science Press, 2002, pages 13-15) and "Protein Technology Handbook" edited by Wang Jiazheng and Fan Ming (Science Press, 2002, pages 60-74), existing Commonly used biomacromolecule concentration methods include: precipitation, adsorption, dialysis, ultrafiltration, vacuum distillation, and freeze-drying. However, the precipitation and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F220/14C08F220/56
Inventor 姚雪彪张颖陈文明窦震
Owner UNIV OF SCI & TECH OF CHINA
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