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Porous integrated substrate of carrying protein solid and its preparation

A protein and overall technology, applied in the direction of carrier binding/immobilization of peptides, peptides, specific peptides, etc., can solve the problems of long time consumption and low group activity, and achieve shortened immobilization time, high reactivity, and improved mass transfer and the effect of convection

Active Publication Date: 2009-06-03
中科榆林能源技术运营有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the overall matrix can be used to immobilize protein molecules, some groups are not highly active, so it often takes a long time to immobilize proteins.

Method used

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  • Porous integrated substrate of carrying protein solid and its preparation
  • Porous integrated substrate of carrying protein solid and its preparation
  • Porous integrated substrate of carrying protein solid and its preparation

Examples

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Embodiment 1

[0034] 1. Preparation of monolithic matrix: press figure 1 As shown, acryloylsuccinimide (NAS) and acrylamide were selected as monomers, and ethylidene dimethacrylic acid (EDMA) was used as a cross-linking agent to synthesize a porous monolithic matrix. The inner wall of the capillary is pre-modified by a silane reagent (3-trimethoxysilyloxypropyl methacrylate, γ-MAPS) with a double bond functional group, and monomers are polymerized in it and fixed on the capillary wall by chemical bonds. Succinimide on NAS is used as an active reactive group to derivatize and immobilize protein molecules. Specifically, such a porous material is polymerized in-situ in a capillary to form a monolithic column and applied to substrate hydrolysis experiments. Weigh 62.5mg of NAS, 62.5mg of acrylamide and 125mg of EDMA, dissolve in 750mg of cyclohexanol / dodecanol (mass ratio 85 / 15) mixed solvent, heat to 60°C, oscillate and mix, cool to 25°C , add 2.5mg of initiator AIBN, after passing nitrogen f...

Embodiment 2

[0041] 1. Preparation of monolithic matrix: press figure 1 As shown, acryloylsuccinimide (NAS) and acrylamide were selected as monomers, and ethylidene dimethacrylic acid (EDMA) was used as a cross-linking agent to synthesize a porous monolithic matrix. The inner wall of the capillary is pre-modified by a silane reagent (3-trimethoxysilyloxypropyl methacrylate, γ-MAPS) with a double bond functional group, and monomers are polymerized in it and fixed on the capillary wall by chemical bonds. Succinimide on NAS is used as an active reactive group to derivatize and immobilize protein molecules. Specifically, such a porous material is polymerized in-situ in a capillary to form a monolithic column and applied to substrate hydrolysis experiments. Weigh and weigh the monomer and porogen in the ratio shown in Table 2, heat to 80°C, oscillate and mix, cool to 25°C, add initiator AIBN, blow nitrogen for 1min, and ultrasonically degas for 5min. At room temperature, inject the above solut...

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Abstract

A protein-carried porous substrate is prepared by taking acryloyl pyrrolidine dione, acrylic amide and ethylene dimethyl acrylic acid as monomers, and in-situ polymerizing in binary high-level alcohol mixed solution to obtain the final product. Protein or pankrin molecules are combined onto integrated substrate tightly. It's simple, stable and fast and has reactive group with high activity.

Description

technical field [0001] The invention relates to a novel porous integral matrix for immobilizing protein and its preparation, which can be used for fast immobilizing protein molecules. Background technique [0002] At present, artificially synthesized matrices for protein immobilization are mainly divided into two categories: granular matrices and monolithic matrices. The preparation of granular matrix generally includes two steps of synthesis and crushing. The obtained particles have different functional groups according to their surface, and generally need to be modified and activated when protein molecules are immobilized. These matrices are mainly used for column packing. In a packed column, the liquid flow mainly passes through the gaps between the fillers, and the mass transfer from the mobile phase to the immobilized enzyme and from the immobilized enzyme to the mobile phase is mainly determined by the diffusion rate, which depends on the filler, the size of the pore...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F220/52C08F220/06C08F2/14C07K17/08
Inventor 张玉奎段继诚张丽华张维冰梁振
Owner 中科榆林能源技术运营有限责任公司
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