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A method for protecting functional protein before ionizing radiation sterilization and its special protective agent

An ionizing radiation and functional technology, applied in the biological field, can solve the problems of cumbersome operation and inability to reduce indirect damage, and achieve the effects of simple operation, protection of functional protein activity, and high safety

Active Publication Date: 2019-02-26
YANTAI ZHENGHAI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the method of Qi Hongzhao and others not only reduces the damage to microorganisms and viruses, but also cannot reduce the indirect damage caused by radiation to BMP-2 activity, and the operation is cumbersome and large-scale production

Method used

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  • A method for protecting functional protein before ionizing radiation sterilization and its special protective agent
  • A method for protecting functional protein before ionizing radiation sterilization and its special protective agent
  • A method for protecting functional protein before ionizing radiation sterilization and its special protective agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Embodiment 1, the preparation of rhBMP-2 stoste

[0085] 1. Construction of recombinant bacteria

[0086] 1. Insert the double-stranded DNA molecule shown in the sequence 1 of the sequence listing from the 64th to the 477th nucleotide at the 5' end between the NdeI and XhoI restriction sites of the vector pET-28a (+) to obtain a recombinant plasmid. In the recombinant plasmid, the inserted double-stranded DNA molecule is fused with part of the DNA of the plasmid itself to form a fusion gene shown in sequence 1 of the sequence listing, and express a fusion protein shown in sequence 2 of the sequence listing.

[0087] 2. Introduce the recombinant plasmid obtained in step 1 into Escherichia coli BL21(DE3) to obtain recombinant bacteria.

[0088] 2. Preparation of rhBMP-2 stock solution

[0089] Take step 1 to obtain the recombinant bacteria, and then follow the purification production method disclosed in the patent CN201310432794.4 through the steps of dissolution of inc...

Embodiment 2

[0092] Example 2, the protective effect of different functional protein protective agents on rhBMP-2

[0093] BMP-2 dimers have the biological activity of inducing osteoblast formation, while BMP-2 multimers and monomers do not have the biological activity. BMP-2 usually needs to be sterilized by irradiation, but cobalt-60γ-ray radiation will produce BMP-2 multimers, thereby losing biological activity. In this embodiment, the purity of BMP-2 dimer in BMP-2 before and after sterilization is detected by high performance liquid chromatography, and the biological activity of BMP-2 before and after sterilization is detected by C2C12 cell method at the same time, so as to evaluate the pretreatment provided by the present invention. The effect of the treatment method.

[0094] 1. Production of samples

[0095] 1. Blank sample

[0096] Get 1 mL of the rhBMP-2 stock solution prepared in Example 1 (the solvent is water, the solute and its concentration are as follows: rhBMP-2 protein...

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Abstract

The invention discloses a method for protecting functional protein or a composite containing functional protein before ionizing radiation sterilization. The method includes the following steps that in a liquid phase system, functional protein or the composite containing functional protein is mixed with a component I and a component II, wherein the component I is carbohydrate, and the component II is amino acid and / or amino acid hydrochloride. As proved by experiments, the method has the effects of resisting radiation and oxidation and protecting the activity of functional protein and has the advantages that all the substances are medicinal auxiliaries and are free of residual toxicity, so that safety is high; the substances of carbohydrate and amino acid are stable in nature and not prone to oxidation, so that stability is high; the method is easy to implement, applicable to experimental sample manufacturing and also suitable for large-scale production.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a method for protecting functional proteins before ionizing radiation sterilization and a special protective agent thereof. The technical problem expected to be solved is to reduce the damage to the activity of functional proteins caused by ionizing radiation sterilization. Background technique [0002] Bone morphogenetic protein (BMP) is a hydrophobic protein secreted by bone matrix and belongs to the transforming growth factor (TGF-β) superfamily member. It was first proposed by Urist et al. A functional protein isolated from matrix extracts. The BMP family includes BMP-1, BMP-2, BMP-4, BMP-6, BMP-7, BMP-9, BMP-12, BMP-14, etc., wherein BMP-2 has the ability to induce undifferentiated fibroblasts to differentiate into The role of osteoblasts, and can regulate the differentiation of osteoblasts and chondrocytes, induce ectopic bone formation and promote fracture healing....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L2/00
CPCA61L2/0029
Inventor 彭继学张东刚屈长宏苑晓佩陈红蕾南南
Owner YANTAI ZHENGHAI BIO TECH
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