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Thiophilic adsorption chromatography monolithic material as well as preparation method and application thereof

A monolithic material and chromatographic technology, applied in the field of thiophilic chromatographic monolithic materials and their preparation, can solve the problems of loss of functionalized groups, inability to reuse multiple times, and high cost

Active Publication Date: 2016-04-06
山东惠分仪器有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the preparation of thiophilic chromatographic materials for enrichment and separation of disulfide compounds is mainly carried out on soft matrices (such as fibers and agarose) or magnetic beads [see J.Chromatogr.B, 2004, 808, 51–56. ; Chromatogr., 2010, 72, 205-213.; Biotech.Progr., 2009, 25, 376-383.] obtained by post-modification of a certain functional group, but these materials have low mechanical strength, high cost, and are prone to loss of functional groups , or cannot be reused multiple times

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  • Thiophilic adsorption chromatography monolithic material as well as preparation method and application thereof
  • Thiophilic adsorption chromatography monolithic material as well as preparation method and application thereof
  • Thiophilic adsorption chromatography monolithic material as well as preparation method and application thereof

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preparation example Construction

[0031] In the preparation method of the present invention, the specific amount of each raw material can be selected in a wide range, but in order to improve the rate and yield of the ring-opening copolymerization reaction, preferably, relative to 1 weight part of TEPIC, the amount of SAA is 0.4 -0.8 parts by weight, the amount of porogen is 2.5-3.2 parts by weight.

[0032] In the preparation method of the present invention, the specific reaction conditions of the ring-opening copolymerization reaction can be selected in a wide range, but in order to improve the rate and the productivity of the ring-opening copolymerization reaction, preferably, the ring-opening copolymerization reaction at least meets the following conditions: Reaction The temperature is 80-150°C, and the reaction time is 2-24h.

[0033] In the present invention, in order to remove unreacted small molecular substances attached to the copolymer poly(TEPIC-co-SAA), thereby improving the purity of the copolymer ...

Embodiment 1

[0046] 1) Dissolve 0.950g of TEPIC in 2.600g of PEG-300, then add 0.550g of SAA into a 10mL centrifuge tube, vortex, ultrasonically degas, and mix evenly to obtain a uniform and clear solution. Block material M1 was obtained by constant temperature copolymerization ring-opening reaction in an oven at 120°C for 12 hours;

[0047] 2) Cut 3.000g of the block material M1 into small pieces, put it into a Soxhlet extractor, add 50mL of methanol, extract at 100°C for 24 hours, wash out the residual porogen and unreacted substances, and obtain the block material M2 .

[0048] 3) Put the bulk material M2 in a vacuum drying oven and dry it at 100°C for 12 hours to obtain the monolithic material A1 of thiophilic chromatography, that is, the copolymer poly(TEPIC-co-SAA).

Embodiment 2

[0050] 1) Rinse the capillary with NaOH (0.1M), water, HCl (0.1M), water and methanol for 30min in sequence; then blow the inner wall of the washed capillary with nitrogen flow for 10h to dry; then at 50°C, dry the The capillary was placed in a mixed solution of ATMS and THF (ATMS / THF v / v 1:1) for contact reaction for 12 hours. After the reaction, the capillary was rinsed with methanol and dried with nitrogen to obtain a capillary with amino groups on the inner wall.

[0051] 2) Completely dissolve 0.095g of TEPIC in 0.260g of PEG-300, then add 0.055g of SAA into a 1.5mL centrifuge tube, vortex and sonicate to obtain a uniform and clear solution. Then, the solution was quickly injected into the capillary of the above-mentioned derivatized amino group, both ends of the capillary were sealed with silicone rubber, and the ring-opening reaction was carried out in a constant temperature oven at 120° C. for 12 hours.

[0052] 3) Take out the above-mentioned capillary tube, connect i...

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Abstract

The invention discloses a thiophilic adsorption chromatography monolithic material as well as a preparation method and an application thereof. The preparation method comprises the following steps: TEPIC adopting a structure shown in a formula (I) and SAA (sulfanilic acid) have ring-opening copolymerization reaction in a pore-foaming agent, and a copolymer poly(TEPIC-co-SAA) is prepared through one step. The thiophilic adsorption chromatography monolithic material prepared with the method has a 3D skeleton structure and is provided with high-density phenolic thioether groups on the surface; meanwhile, the method adopts a one-step method, the yield is further increased and the operation process is simplified; besides, the thiophilic adsorption chromatography monolithic material has excellent effects on enrichment and separation of disulfide.

Description

technical field [0001] The present invention relates to a thiophilic material, in particular to a thiophilic chromatographic monolithic material and its preparation method and application. Background technique [0002] Thiophilic chromatography, also known as thiophilic chromatography, is a simple and economical protein purification technique, especially for the development of antibody purification methods. Thiophilic chromatography produces specific adsorption on antibodies and certain other proteins in a high-salt environment, and then elutes at a low salt concentration, so that high-purity and high-recovery protein products can be obtained. It is a new protein purification technology. [0003] Due to the active epoxy groups in the molecular structure of epoxides, they can react with various types of materials to form high polymers with a network structure. Among them, the molecular structure of epoxy resin is dense, and the groups contained in its curing system are highl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/282B01J20/30B01D15/20C07K1/22C07K16/00
Inventor 刘云春李晓青
Owner 山东惠分仪器有限公司
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