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Preparation method of recombinant human growth hormone and preparation method of PEGylation modification substance

A technology for the modification of human growth hormone, which is applied in the field of preparation of polyethylene glycol modifications, can solve the problems of foreign protein interference, complex process steps, low activity of human growth hormone, etc., and achieve rapid and efficient separation and high application value Effect

Inactive Publication Date: 2015-09-30
杭州北斗生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Practice has proved that the production of secreted recombinant human growth hormone, although the steps are simple, the interference of foreign proteins is less, and the environment of the oxidized type is conducive to protein folding, but its expression level is very low, which is generally more than 1 order of magnitude lower than that of the intracellular type
However, the production of intracellular proteins has disadvantages such as complex process steps and more interference from foreign proteins.
In addition, the human growth hormone obtained by the above two methods also has defects such as low activity and low purity.

Method used

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  • Preparation method of recombinant human growth hormone and preparation method of PEGylation modification substance
  • Preparation method of recombinant human growth hormone and preparation method of PEGylation modification substance
  • Preparation method of recombinant human growth hormone and preparation method of PEGylation modification substance

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Experimental program
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Effect test

Embodiment 1

[0054] The recombinant human growth hormone used in this example has a purity greater than 95% and maintains high biological activity.

[0055] Dissolve recombinant human growth hormone (rhGH) in 100Mm phosphate buffer (PH10), add PEG amber according to the ratio of recombinant human growth hormone (rhGH): polyethylene glycol (PEG) = 1:3 (W / W) Imido-succinamide ester (mPEG-SS), stirred and dissolved, reacted at 4°C for 2 hours.

Embodiment 2

[0057] The recombinant human growth hormone used in this example has a purity greater than 95% and maintains high biological activity.

[0058] Dissolve recombinant human growth hormone (rhGH) in 100Mm phosphate buffer (PH8), add PEG amber according to the ratio of recombinant human growth hormone (rhGH): polyethylene glycol (PEG) = 1:6 (W / W) Imido-succinamide ester (mPEG-SS), stirred and dissolved, reacted at 4°C for 2 hours.

Embodiment 3

[0061] The product obtained by reacting recombinant human growth hormone (rhGH) with polyethylene glycol was dissolved in 100Mm phosphate buffer (PH8) and dialyzed, after being adsorbed by anion exchange resin or cation exchange resin, it was eluted with salt, and polyethylene glycol was collected. The alcoholized recombinant human growth hormone drug is concentrated and purified by molecular sieve gel chromatography. The purity of the obtained polyethylene glycol recombinant human growth hormone drug can be greater than 95%, and the product is obtained after small filtration and freeze-drying .

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Abstract

The invention relates to a gene recombinant human growth hormone modification substance for treating microsoma caused by children's endogenous recombinant human growth hormone deficiency or insufficiency and for surgeries for injuries and serious burns and subsequent treatment, particularly a preparation method of a polyethyleneglycol (PEG) modification substance of the growth hormone. The invention aims to provide a preparation method of a recombinant human growth hormone, which is higher in expression level and convenient for operation, can enhance the purity and activity of the effective drug and can enhance the purity and activity of the product on the premise of not increasing the cost, thereby enhancing the activity of the drug. The invention also aims to provide a preparation method of the recombinant human growth hormone PEGylation modification substance. According to the method, the PEGylation is utilized to modify the drug, so that the PEG and human growth hormone are connected through covalence, thereby prolonging the half life of the drug and lowering the toxic or side effect of the drug protein.

Description

technical field [0001] The present invention relates to a gene recombinant human growth hormone modifier for treating short stature caused by lack or deficiency of endogenous recombinant human growth hormone in children and for operation and follow-up treatment of injuries and severe burns, in particular to the polymorphism of growth hormone Preparation method of ethylene glycol modified product. Background technique [0002] Human growth hormone (human growth th ho rmone, hGH) is an important hormone secreted by eosinophilic cells in the anterior pituitary gland, which has biological functions of promoting human growth and development and maintaining tissue and organ functions. Recombinant human growth hormone (rhGH) is mainly used clinically to treat short stature in children, but also widely used in surgery and follow-up treatment of injuries and severe burns. Human growth hormone (HGH), also known as somatotorpin, is a hydrophilic single chain globulin composed of 191 a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/18C12N15/70C12N1/21C07K14/61C07K1/22C07K1/107
Inventor 封小燕张加慧刘沐荣徐滔李蕾
Owner 杭州北斗生物技术有限公司
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