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A kind of preparation method of protein affinity imprinted hydrogel polymer based on acrylamide group metal chelating monomer

A hydrogel polymer and acrylamide technology, applied in the field of molecularly imprinted polymers, can solve problems such as difficult adsorption and desorption, slow mass transfer process of template molecules, and weak ion exchange force

Active Publication Date: 2020-08-18
NANJING BAISAI BIOCHROMATOGRAPHY TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

2. This research is a surface imprinting method. Although the surface imprinting method can allow macromolecules to freely enter and exit the bonding site and make the surface a specific target site for imprinting proteins, surface imprinting needs to obtain carrier materials through complicated preparation steps. , and then carry out the surface molecular imprinting reaction with the support of the carrier material, the preparation process of this method is cumbersome, the cost is high, and it is not suitable for large-scale industrialization and application
3. The copper ions in this study are combined with the carboxylic acid groups on the polymer in the form of ion exchange, the ion exchange force is weak, the ion exchange capacity is very low, and it is easily lost by the competition of other monovalent cations in the buffer system. Reduce protein adsorption capacity
[0007] Secondly, in terms of the initiation method, for protein molecular imprinting, its structural characteristics limit the need to retain the biological activity of the molecule itself after imprinting, and protein molecules are sensitive to temperature and organic solvents. Generally, molecular imprinting uses organic solvents as the reaction system. Polymerization is usually triggered by heating, resulting in changes in the secondary structure of the protein and loss of activity
[0008] In addition, traditional molecularly imprinted polymers need to rely on a high degree of crosslinking to maintain the rigidity of their imprinted structures. The prepared polymers are highly crosslinked, but too high crosslinking not only makes the external shape of molecularly imprinted polymers loose and brittle , also lead to slow mass transfer process of template molecules in the polymer, difficult adsorption and desorption, and slow desorption of residual template molecules during use

Method used

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  • A kind of preparation method of protein affinity imprinted hydrogel polymer based on acrylamide group metal chelating monomer
  • A kind of preparation method of protein affinity imprinted hydrogel polymer based on acrylamide group metal chelating monomer
  • A kind of preparation method of protein affinity imprinted hydrogel polymer based on acrylamide group metal chelating monomer

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] In a 2L four-necked round-bottomed flask equipped with a stirring paddle, a thermometer, a condenser tube and a feeding port, add 200ml of chloroform, add 50g of ethyl methacrylate and 0.5 mole of biaminodiacetic acid and stir to mix, add 1.8g React with anhydrous potassium hydroxide, slowly raise the temperature to 80°C, and react at this temperature for 3 hours, distill under reduced pressure to remove the unreacted ethyl methacrylate and the reaction product ethanol, and then add 1mol / l NaOH dropwise Adjust the pH to 10 and transfer to a 500ml separatory funnel. Add 100ml of chloroform to the separatory funnel, shake it, let it stand for stratification, remove the upper aqueous solution, and dry it under reduced pressure on a rotary evaporator to obtain 85.3g of a white product, which is the monomer methyl of the acrylamide family that can chelate with metal ions. Sodium Acrylaminodiacetate.

Embodiment 2

[0048] In a 2L four-necked round-bottomed flask equipped with a stirring paddle, a thermometer, a condenser tube and a feeding port, add 200ml of chloroform, 100g of methacryloyl chloride and 0.5 molar ratio of aminotriacetic acid and stir to mix, add 5g of anhydrous hydrogen Potassium oxide was reacted, and the temperature was slowly increased to 60°C. After reacting at this temperature for 6 hours, the unreacted methacryloyl chloride and the reaction product ethanol were distilled off under reduced pressure, and then 1mol / l NaOH was added dropwise to adjust the pH to 10 , transferred to a 500ml separatory funnel. Add 180ml of chloroform to the separatory funnel, shake it, let it stand for stratification, remove the upper aqueous solution, and dry it under reduced pressure on a rotary evaporator to obtain 152.6g of a white product, which is a monomeric methyl compound of the acrylamide family that can be chelated with metal ions. Sodium Acrylaminotriacetate.

Embodiment 3

[0050] (1) Synthesis of bovine serum albumin-imprinted hydrogel polymer (MIP)

[0051] A) In the aqueous medium, add 60 mmoL of sodium methacrylate aminooxalate, a monomer of acrylamide family that can chelate metal ions, add 10 mmoL of nickel sulfate while stirring slowly, and continue stirring for 30 min to form a stable complex. compound;

[0052] B) then add the bovine serum albumin BSA powder of 200mg, continue stirring to make it fully dissolve;

[0053] C) Then add 30 mmol of cross-linking agent N,N'2 methylenebisacrylamide, 100 mg of initiator ammonium persulfate and 20 mg of emulsifier Tween 80.

[0054] D) Molecular imprinting was carried out by reverse-phase suspension cross-linking method, and the above-mentioned aqueous phase imprinting system was prepared into W / O water-in-oil emulsion droplets. The specific method is as follows: add the above aqueous phase solution to the reverse suspension medium composed of chloroform, n-hexane and Span 80, the volume ratio ...

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Abstract

The invention belongs to the technical field of molecularly imprinted polymers, and particularly relates to a preparation method of protein affinity blotting aquagel polymers based on acrylamide groupmetal chelation monomers. The invention aims at providing the preparation method of the protein affinity blotting aquagel polymers based on the acrylamide group metal chelation monomers by aiming atthe technical analysis and the existing problems. The preparation method of the protein affinity blotting aquagel polymers based on the acrylamide group metal chelation monomers has the technical scheme that according to the method, after the acrylamide group monomers are chelated with divalent metal ions, target protein is subjected to specific adsorption combination and removal; then, polymers are obtained. The preparation method has the following characteristics that the acrylamide group monomers have the metal chelation capability; the molecular mark aquagel has the temperature and pH duplex sensitivity; the reverse phase suspension technology is used; the polymers are aquagel polymers.

Description

technical field [0001] The invention belongs to the technical field of molecularly imprinted polymers, in particular to a method for preparing protein affinity imprinted hydrogel polymers based on acryloyl group metal chelating monomers. [0002] technical background [0003] Molecular imprinting technology refers to the technology of preparing polymers with selective recognition ability for a specific molecule (usually called template molecule or imprinted molecule). The prepared polymer is called molecularly imprinted polymer, or MIP for short. MIP is a polymer with high chemical and pH stability, can withstand high temperature, high pressure and harsh acid-base conditions, has a long service life, is simple to prepare, and has three-dimensionally matched holes with template molecular structure and functional units. Specific recognition sites (catalytic or hydrogen bonding) for body energy formation. [0004] Molecularly imprinted polymers have been successfully applied in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F220/58C08F222/38C08F2/32B01J20/26B01J20/30
CPCB01J20/261B01J2220/4812C08F2/32C08F220/58C08F222/385C08F222/38
Inventor 于晓军刘吉众彭琳
Owner NANJING BAISAI BIOCHROMATOGRAPHY TECH
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