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Method for reducing DNA impurities in viral compositions

一种组合物、病毒的技术，应用在病毒纯化,狂犬病病毒的生产,Vero细胞领域，能够解决天然摄入率不高等问题，达到克服局限性的效果

Active Publication Date: 2013-04-17
BEIJING YISHENG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compositions for oral administration are recommended to be less than 100 μg / dose as the natural uptake rate of this method of administration is not high

Method used

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  • Method for reducing DNA impurities in viral compositions
  • Method for reducing DNA impurities in viral compositions
  • Method for reducing DNA impurities in viral compositions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] Example 1 - Old procedure without using high ionic concentration buffers

[0117] This example illustrates how the residual DNA content of a rabies virus solution is not reduced when the purification process does not use buffers of high ionic concentration to wash the virus composition.

[0118] CTN rabies virus was grown in Vero cells and live rabies virus was harvested from the supernatant, ie without the use of nucleases. Viral supernatants were first purified to remove residual DNA from the suspension, then live rabies virus particles were concentrated to up to 50-fold the original concentration by ultrafiltration with low salt concentration prior to chemical inactivation with β-propiolactone buffer rinse. At this point, the antigen content and residual DNA were measured using the standard test method specified in the Chinese Pharmacopoeia 2010 (see Table 1 below).

[0119] Afterwards, the virus solution was subjected to size exclusion chromatography using 4FF aga...

Embodiment 2

[0125] Example 2 - Effect of Ion Concentration on DNA Clearance at Laboratory Scale

[0126] This example illustrates the effect of different ion concentrations on reducing the residual DNA content of a rabies virus solution on a laboratory scale.

[0127] CTN rabies virus was grown in Vero cells and live rabies virus was harvested from the supernatant, ie without the use of nucleases. Viral supernatants were filtered, then concentrated by ultrafiltration to up to 50-fold the original concentration, rinsed in high ionic buffer and then in low salt buffer prior to chemical inactivation with β-propiolactone Rinse in. At this point, the antigen content and residual DNA were measured using the standard test method specified in the Chinese Pharmacopoeia 2010 (see Table 2 below).

[0128] Afterwards, wash the virus solution with high ionic concentration buffer (containing 0.5M (B), 1M (C), 2.0M (D), 3.0M (E) NaCl) and regular PBS buffer (A). After rinsing the virus solution, size...

Embodiment 3

[0134] Example 3 - Novel process for large scale use of high ionic concentration buffers

[0135] This example illustrates how reducing the residual DNA content of a rabies virus solution preserves the immunogenicity of rabies virus particles.

[0136] CTN rabies virus was grown in Vero cells and live rabies virus was harvested from the supernatant, ie without the use of nucleases. Viral supernatants were filtered, then concentrated by ultrafiltration to up to 50-fold the original concentration, rinsed in high ionic buffer and then in low salt buffer prior to chemical inactivation with β-propiolactone Rinse in. At this time, the antigen content and residual DNA were measured with the standard test method specified in the Chinese Pharmacopoeia 2010 (see Table 3 below).

[0137] Then, wash the virus solution with a high ion concentration buffer containing 1M NaCl. After washing, size exclusion chromatography was performed using 4FF agarose medium. At this point, the antigen ...

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Abstract

A method for purification of viral compositions is provided. In particular, a method for reduction of unwanted residual DNA in a viral composition while retaining the immunogenicity of the virus itself is provided. The resulting immunogenic viral composition is substantially free of residual DNA, and is useful for the manufacture of medical products, such as vaccines designed for human or animal.

Description

technical field [0001] The present invention relates generally to the manufacture of vaccines, and in particular to the purification of viruses, and more particularly to the purification of viruses by eliminating residual DNA from the cellular matrix used for virus propagation. In particular, the invention relates to virus production using passaged cell lines, especially Vero cells. The production process may include harvesting the viral particles of interest, purifying the viral composition, inactivating the virus, and mixing the viral composition with a solution with high ionic strength, followed by further purification by size exclusion chromatography. The invention is applicable to the production of viruses, and in particular the production of rabies virus, for the manufacture of medical products such as vaccines for humans and animals. Background technique [0002] 1 Introduction [0003] Engineered virus particles can be used in a variety of human medical application...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K39/205A61P31/14C12N7/02C12R1/93
CPCA61K2039/525C12N2760/20134C12N2760/20151A61K39/12C12N7/00A61P31/14C12N7/02C12N7/06
Inventor 张译代金明倪亚瑾
Owner BEIJING YISHENG BIOTECHNOLOGY CO LTD
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