Efficient protective reagent for virus purification and application of efficient protective reagent

A technology for protecting reagents and viruses, applied in the field of biopharmaceuticals, can solve problems affecting the purification yield of solid AAV, inactivation of biologically active solid AAV, etc., and achieve the effects of avoiding oxidative denaturation, preventing aggregation, and stabilizing biofilm and protein structure

Pending Publication Date: 2022-04-15
苏州博腾生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The first object of the present invention is to aim at the prior art described in the background technology, when affinity chromatography is carried out to adeno-associated virus (AAV), the biological activity that affects AAV and when removing empty shell AAV will also make The inactivation of solid AAV affects the purification yield of solid AAV, and provides a virus purification and high-efficiency protection reagent that can solve the aforementioned problems

Method used

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  • Efficient protective reagent for virus purification and application of efficient protective reagent

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Embodiment 1

[0014] A virus purification and high-efficiency protection reagent, which includes components and contents: 1 sucrose, 30g / L arginine, 20g / L glycine, 20g / L mannitol, 40g / L trehalose, 0.5M MgCl 2 , 0.5M Tris-HCl, and the rest is ultrapure water with a pH of 7.5.

Embodiment 2

[0016] A virus purification and high-efficiency protection reagent, which includes components and contents: 0.5 sucrose, 20g / L arginine, 15g / L glycine, 15g / L mannitol, 35g / L trehalose, 0.4M MgCl 2 , 0.5M Tris-HCl, and the rest is ultrapure water with a pH of 7.5.

Embodiment 3

[0018] A virus purification and high-efficiency protection reagent, which includes components and contents: 0.8 sucrose, 25g / L arginine, 18g / L glycine, 18g / L mannitol, 38g / L trehalose, 0.5M MgCl 2 , 0.5M Tris-HCl, and the rest is ultrapure water with a pH of 7.5.

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Abstract

The invention discloses a virus purification efficient protection reagent and application thereof, the virus purification efficient protection reagent comprises the following components: 0.5-1 M of sucrose, 20-30 g / L of arginine, 15-20 g / L of glycine, 15-20 g / L of mannitol, 35-40 g / L of trehalose, 0.4-0.5 M of MgCl2, 0.5 M of Tris-HCl, and the balance of ultrapure water, and the pH value is 7.5. According to the efficient protection reagent for virus purification, AAV viruses or other viruses can be protected by adding the efficient protection reagent for virus purification in a balancing stage, a sample loading stage and an elution stage in a virus chromatographic purification process; the influence on the activity and yield of the AAV or other viruses due to protein conformation change caused by dramatic change of conditions in chromatographic purification of protein shells of the AAV or other viruses is avoided.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, in particular to a virus purification and high-efficiency protection reagent. Background technique [0002] There are mainly two methods for virus purification in the prior art, namely density gradient centrifugation and chromatographic purification. Density gradient centrifugation mostly uses cesium chloride or iodixanol for density gradient centrifugation, but gradient centrifugation faces many difficulties such as high cost and long time consumption when used in pilot scale-up and GMP production. In the chromatographic purification method, affinity chromatography and ion chromatography are mostly used. Excessively high or low ion concentration in ion chromatography will affect the charge of AAV, affect the folding of AAV protein, lead to conformational changes, and even cause protein denaturation and loss of activity. [0003] When performing affinity chromatography on adeno-assoc...

Claims

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Application Information

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IPC IPC(8): C12N7/02C12R1/93
CPCC12N7/00C12N2750/14151
Inventor 张海峰王雷刘佳博隋礼丽钱浩
Owner 苏州博腾生物制药有限公司
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