Long-term storage method for proteins

A long-term preservation, protein technology, applied in the field of biomedicine, can solve problems such as hindering application, and achieve the effects of controllable parameters, good reproducibility, and little structural impact

Active Publication Date: 2021-08-13
康汉医药(广州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The shortcoming of this thermal instability of most proteins is one of the important factors hindering its application in biomedicine, enzyme engineering, food industry, etc. Therefore, the improvement of protein thermal instability has always been a focus and difficulty

Method used

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  • Long-term storage method for proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The preparation of embodiment 1 amphiphilic polymer

[0040] Prepare amphiphilic polymers as follows:

[0041] S01. Dissolving the hydrophobic polymer material in an organic solvent, adding a hydrophilic polymer material for a mixed reaction, removing the organic solvent and other impurities in the reaction system to obtain a mixture A;

[0042] S02. The mixture A was redissolved in water, and the product was collected after purification.

[0043] S03. Collect the dialysis product, freeze overnight at -80°C, and then freeze-dry on a freeze dryer to obtain a white flocculent substance.

[0044]Hydrophilic polymer materials include polyethylene glycol, polyoxyethylene, mPEG-NH2-400, mPEG-NH2-600, mPEG-NH2-800, mPEG-NH2-1000, mPEG-NH2-2000, mPEG-NH2- 4000, mPEG-NH2-5000 or mPEG-NH 2 -At least one in 10000; Select polyethylene glycol for use in the present embodiment.

[0045] The hydrophobic polymer material preferably comprises polymaleic anhydride, poly(maleic anhydr...

Embodiment 2

[0053] Example 2 Preparation of Amphiphilic Polymer and Detection of Self-Assembly

[0054] Follow the steps below to prepare amphiphilic polymers

[0055] S01. Dissolve poly(maleic anhydride-ALT-1-octadecene) in organic solvent, add mPEG-NH in proportion 2 Mixing reaction, removing the organic solvent and other impurities in the reaction system, to obtain mixture A;

[0056] S02. Reconstitute the mixture A with water, put it into a dialysis bag and perform dialysis in pure water;

[0057] S03. Collect the dialysis product, freeze overnight at -80°C, and then freeze-dry on a freeze dryer to obtain a white flocculent substance.

[0058] The organic solvent generally selects dichloromethane, dimethyl sulfoxide, acetone, ethanol, and selects dichloromethane in this embodiment.

[0059] mPEG-NH 2 Grafting ratio P with poly(maleic anhydride-ALT-1-octadecene) m m n (1:10)~(2:1), respectively prepare P 1 m 10 ,P 1 m 5 ,P 2 m 1 ,P 1 m 1 ,P 2 m 1 Five amphiphilic polyme...

Embodiment 3

[0065] The method for the long-term preservation of embodiment 3 bovine serum albumin

[0066] (1) Bovine serum albumin is dissolved in water to obtain bovine serum albumin aqueous solution; the amphiphilic polymer P described in step (1) 1 m 10 and a cosolvent (DMSO) were dissolved in water to obtain an amphiphilic polymer suspension.

[0067] (2) Add the amphiphilic polymer suspension described in step (1) dropwise into the bovine serum albumin aqueous solution according to the mass ratio of 2:1, stir on a magnetic stirrer until clear, and the stirring speed is about 400rpm, Stir for about 2h.

[0068] (3) Perform dialysis and purification on the protein-polymer mixed solution obtained in step (2), select a dialysis bag corresponding to the molecular weight cut-off, remove the protein not bound to the polymer, and obtain the high heat stable bovine serum albumin.

[0069] The high temperature resistance ability of the prepared high heat stable bovine serum albumin was det...

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Abstract

The invention discloses a long-term storage method for proteins. The method comprises the steps of dissolving a protein drug, then, adding an amphiphilic polymer suspension, carrying out uniform mixing thoroughly, and carrying out purifying, thereby obtaining a polymer-protecting protein drug solution. According to the method, the original protein structure of the protein drug is not destroyed, the drug effect of the protein drug will not be hindered, and thus, the ubiquitous problem of proteins that the proteins are short in storage time and are prone to denaturation and deterioration at normal temperature is excellently solved. The method can be used for universally improving the thermal stability of the protein drug, the thermal stability of the protein drug is improved remarkably, the application of the protein drug in the aspect of biomedicines is extended, and thus, the method has a broad application prospect.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and more specifically relates to a method for improving the thermal stability of protein drugs. Background technique [0002] In terms of biomedicine, protein drugs (vaccines, hormones, cytokines, etc.) generally have the problem of short storage period at room temperature, and usually require cold chain protection during storage and transportation, which leads to their higher market prices, improving these The thermal stability of drugs will expand its application in developing countries; in enzyme engineering, proteins, as biocatalysts, often need to withstand high temperature reaction conditions, if their stability at high temperatures can be improved, the reaction will be greatly improved. Efficiency, so that the green, economical and sustainable industrial production method of enzyme catalysis is more widely used; in the food industry, food protein is inevitably affected by heat during p...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C07K14/765C07K14/805C07K14/79C08G81/02
CPCC07K14/765C07K14/805C07K14/79C08G81/025
Inventor刘小文
Owner康汉医药(广州)有限公司