Biomolecule immobilization method and application thereof

A technology of biomolecules and reaction solvents, applied in biochemical equipment and methods, immobilized enzymes, chemical instruments and methods, etc., can solve the problem of low stability of biomolecules, reduce storage difficulty and cost, and maintain biological activity. , the effect of good economic benefits

Pending Publication Date: 2022-08-02
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to overcome the problem of low stability of biomolecules in the prior art, and to provide a method for immobilizing biomolecules and its application. This method can quickly immobilize biomolecules and improve the biomolecules. Stable, and efficiently maintain the biological activity of biomolecules

Method used

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  • Biomolecule immobilization method and application thereof
  • Biomolecule immobilization method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0039] 5 mg of cytochrome c was added to 4.5 mL of deionized aqueous solution containing 2-methylimidazole, sodium caseinate, and polyvinylpyrrolidone (2-methylimidazole concentration was 160 mM, sodium caseinate concentration was 1.2 mg / mL, The concentration of polyvinylpyrrolidone is 1 mg / mL, pH=10.3), and then mixed with 0.5 mL of deionized aqueous solution containing zinc acetate (the concentration of zinc acetate is 40 mM), and placed in contact for 10 min at pH 10 and room temperature to obtain a reaction solution. The reaction solution was centrifuged at 6000 rpm for 10 min, the precipitated product was collected, and the precipitated product was washed twice with deionized water to obtain immobilized cytochrome c.

Embodiment 2

[0041] 5 mg of cytochrome c was added to 2.5 mL of deionized aqueous solution containing 2-imidazole carboxaldehyde, sodium caseinate, and polyvinylpyrrolidone (2-methylimidazole concentration was 400 mM, sodium caseinate concentration was 1.2 mg / mL, poly Vinylpyrrolidone concentration of 0.9mg / mL), then mixed with 2.5mL of deionized aqueous solution containing 74.26mg of zinc acetate, placed in contact at pH 8 and room temperature for 5min to obtain a reaction solution, and the reaction solution was centrifuged at 5000rpm. After 12 min, the precipitated product was collected, and the precipitated product was washed twice with deionized water to obtain immobilized cytochrome c.

Embodiment 3

[0043] 5 mg of glucose oxidase was added to 5 mL of deionized aqueous solution containing 2-methylimidazole, sodium caseinate, and polyvinylpyrrolidone (2-methylimidazole concentration was 160 mM, sodium caseinate concentration was 1.5 mg / mL, poly Vinylpyrrolidone concentration of 1.5mg / mL), then mixed with 0.75mL of deionized aqueous solution containing zinc acetate (zinc acetate concentration of 40mM), placed in contact at pH 12 and room temperature for 15min to obtain a reaction solution, the reaction solution was rotated at the rotating speed Centrifuge at 8000 rpm for 8 min, collect the precipitated product, and wash the precipitated product twice with deionized water to obtain immobilized glucose oxidase.

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Abstract

The invention relates to a biological immobilization technology, and discloses a biomolecule immobilization method and application thereof. The immobilization method of the biomolecules comprises the following steps: in the presence of a reaction solvent, contacting the biomolecules, a metal salt and a forming agent, wherein the biomolecules are cytochrome c and/or glucose oxidase, and the forming agent contains imidazole and/or imidazole derivatives. The invention also provides the immobilized biomolecule obtained by the immobilization method, and an application of the immobilized biomolecule in at least one of a biological detector, food production and preparation of a clinical injection. According to the method, the biomolecules can be rapidly immobilized, the stability of the biomolecules is improved, and the biological activity of the biomolecules is efficiently maintained.

Description

technical field [0001] The present invention relates to bioimmobilization technology, in particular, to a method for immobilizing biomolecules and its application. Background technique [0002] Cytochrome c (Cyt c) is a soluble pigment protein with iron porphyrin as a prosthetic group, and is one of the main members of the cellular respiratory chain. The outer surface of the mitochondrial membrane within the cardiac muscle cytoplasm. Cytochrome c is a very important electron transporter in the process of biological oxidation, which accelerates the enzymatic action, and has a very good curative effect on paralysis, emphysema and other diseases caused by tissue hypoxia, especially when the disease deteriorates and rescues , Intravenous injection of this product has a good effect, play a role in adjusting cellular respiration and material metabolism. However, cytochrome c is sensitive to the environment and weak in stability, which greatly limits its application range. [00...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/02C12N11/082C12N9/04C07K17/02C07K17/08C07K14/80
CPCC12N11/02C12N11/082C12N9/0006C12Y101/03004C07K17/02C07K17/08C07K14/80
Inventor 黄和张幸周青倩陈瑶
Owner NANJING NORMAL UNIVERSITY
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