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A kind of monolithic material of phospholipid organic polymer and its preparation method and application

A technology of monolithic materials and polymers, applied in the field of protein purification, can solve the problems of incompatibility between specificity and recovery, lack of specificity, reproducibility, and purity to be further verified

Active Publication Date: 2020-10-27
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can achieve the effect of one-step purification of CRP in human serum to a certain extent, but there are still bands of other foreign proteins in the purified solution, and the purity needs to be further verified; and this method does not carry out specificity and reproducibility Such investigations are not enough for subsequent promotion [Eunjoo Kim, Se Geun Lee, Hyun-ChulKim et al. Separation Science and Technology, 2013, 48: 2600-2607]
Therefore, the current commercial CRP purification technology still uses agarose gel as the carrier and phospholipid materials as the ligand (Immobilized p-Aminophenyl Phosphoryl Choline Gel). Shown [L. Soler, N. García, A. Unzueta et al. Vet Immunol Immunop, 2016, 179:26–31; Michael G. Roper, Megan L. Frisk, Janeen P. Oberlander. Anal Chim Acta, 2006, 569:195–202]

Method used

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  • A kind of monolithic material of phospholipid organic polymer and its preparation method and application
  • A kind of monolithic material of phospholipid organic polymer and its preparation method and application
  • A kind of monolithic material of phospholipid organic polymer and its preparation method and application

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Experimental program
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Effect test

Embodiment 1

[0037] Phospholipid monomer compound 2-methacryloyloxyethylphosphorylcholine (2-methacryloyloxyethylphosphorylcholine, MPC), cross-linking agent MBA, porogen (mixed system of IPA and THF) and initiator AIBN, according to the optimal ratio Prepare a polymerization reaction mixed solution, ultrasonically dissolve, degas, and pour into a 200 μL pipette tip, then seal both ends of the pipette tip, and put it in a 60°C water bath for 12 hours; after the reaction, put the pipette tip with The syringe was connected, and the porogen, unreacted monomer, cross-linking agent and oligomer were washed away with methanol on the peristaltic pump to obtain a phospholipid organic polymer monolithic column (poly(MPC-co-MBA)). Wherein the mass ratio of monomer MPC and porogen is 25:75, the mass ratio of monomer MPC and crosslinking agent MBA is 70:30, the mass ratio of porogen IPA and THF is 60:40, and the mass ratio of initiator AIBN The mass is 1% of the monomeric MPC.

[0038] The scanning e...

Embodiment 2

[0040] The phospholipid monomer compound MPC, the cross-linking agent PDA, the porogen (a mixed system of DMSO and THF) and the initiator AIBN were formulated into a polymerization reaction mixed solution according to the optimal ratio, ultrasonically dissolved, degassed, and poured into a 200 μL pipette Then seal both ends of the pipette tip and put it in a 60°C water bath to react for 12 hours; after the reaction is completed, connect the pipette tip to the syringe, and rinse off the porogen and unreacted porogen with methanol on the peristaltic pump. Monomers, cross-linking agents and oligomers were used to obtain poly(MPC-co-PDA) organic polymer monoliths. Wherein the mass ratio of monomer MPC and porogen is 15:85, the mass ratio of monomer MPC and crosslinking agent PDA is 80:20, the mass ratio of DMSO and THF is 40:60 in the porogen, and the mass ratio of initiator The mass is 1% of the total amount of monomeric MPC.

Embodiment 3

[0042] The phospholipid monomer compound MPC, the cross-linking agent EDMA, the porogen (a mixed system of IPA and BDO) and the initiator AIBN were formulated into a polymerization reaction mixed solution according to the optimal ratio, dissolved by ultrasonic, degassed, and poured into a 200 μL pipette Then seal both ends of the pipette tip and put it in a 60°C water bath for 12 hours of reaction; porogens and oligomers to obtain poly(MPC-co-EDMA) organic polymer monoliths. Wherein the mass ratio of monomer MPC and porogen is 15:85, the mass ratio of monomer MPC and crosslinking agent EDMA is 53:47, the mass ratio of IPA and BDO is 25:75 in the porogen, and the mass ratio of initiator The mass is 1% of the mass of monomeric MPC.

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Abstract

The invention belongs to the technical field of protein purification, and discloses a phospholipid organic polymer whole material and a preparation method and application thereof. The method comprisesthe steps that phospholipid monomeric compound, crosslinking agent, porogen and initiator are mixed and then poured into a container after ultrasonic dissolving and deaeration are conducted, aggregation reaction is conducted under the temperature of 40-70 DEG C or under ultraviolet irradiation, and the phospholipid organic polymer whole material is obtained. The phospholipid organic polymer wholematerial has the advantages of high reproducibility, good biocompatibility and protein nonspecific adsorption resistance, and the existing defects of a traditional CRP purified material are effectively overcome. According to the material, by taking the phospholipid organic polymer whole material as an absorbent purifying standard protein mixed sample or an actual sample, CRP proteins with high purity can be obtained simply by one purifying strategy, the operation is easy, and the recovery rate is close to 100%.

Description

technical field [0001] The invention belongs to the technical field of protein purification, and in particular relates to a phospholipid organic polymer integral material, a preparation method and application thereof. Background technique [0002] In 1930, Tillett and Francis discovered that in patients with pneumonia, there is a substance that can react with the phospholipid components on the cell wall of Streptococcus pneumoniae C polysaccharide (CPS) to produce a precipitate [W.S.Tillett,, T.Jr.Francis, J.Exp .Med.,1930,52:561], later called C-reactive protein (CRP). Native CRP is a non-covalent pentameric structure composed of 5 independent subunits, produced by hepatocytes, and mainly present in serum, ascites and cerebrospinal fluid [J.P.Atkinson,Arthritis Rheum.,2001,44:995– 996.], its content is very low in the normal body, and its content will increase hundreds of times during inflammatory reactions, so it has become a non-specific acute phase protein [M.McCarty, J...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F230/02C08F222/38C08F222/14C08F130/02C08J9/28C07K14/47C07K1/22
CPCC07K14/4737C08F130/02C08F230/02C08J9/28C08J2343/02C08F222/102
Inventor 江正瑾王启钦金含颖夏东海邵慧凯王祥宇
Owner JINAN UNIVERSITY
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