Verification method of low pH incubation virus inactivation

A verification method and virus technology, applied in the biological field, can solve the problems of strict specifications of the verification experiment of pH incubation inactivated virus, and achieve the effects of optimizing technical details, improving titer and stable quality.

Inactive Publication Date: 2018-07-31
CANVEST WUHAN BIOTECH
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Problems solved by technology

However, these two guidance documents did not make strict specifications on the technical details of the verification experiment of low pH ...

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  • Verification method of low pH incubation virus inactivation
  • Verification method of low pH incubation virus inactivation
  • Verification method of low pH incubation virus inactivation

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[0049] The following examples will further illustrate other features and advantages of the present invention, but these examples are only for illustration, not limitation of the present invention.

[0050] All materials and reagents used below are commercially available unless otherwise specified.

[0051] The present invention provides a kind of verification method of low pH hatching inactivated virus, and the specific technical scheme that it takes is as follows:

[0052] 1. Amplification, purification and titer determination of indicator virus

[0053] Pseudorabies virus (PRV), murine leukemia virus (MuLV), vesicular stomatitis virus (VSV), and porcine parvovirus (PPV) were selected as indicator viruses (all purchased from China Type Culture Collection Center). (CCTCC)). Its virus properties are shown in Table 1, covering DNA viruses of various genera and hosts, RNA viruses, enveloped viruses and non-enveloped viruses.

[0054] Table 1. Indicator viruses in the low pH in...

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Abstract

The invention relates to a verification method of low pH incubation virus inactivation. The method comprises the steps of selecting indicator viruses and corresponding host cells; amplifying and purifying the indicator viruses; titering virus working primary liquid and samples; optimizing the pH value of virus inactivation verification experiments; estimating the low pH incubation virus inactivation effect. By means of the method, the full process of genetically engineered drug production technology virus inactivation can be simulated in a laboratory, and the amplifying and purifying technology of the indicator viruses is optimized, so that the titer of the working primary liquid of the indicator viruses is increased, the pH value is unified to 7.0, the quality is stable, and subsequent operation is facilitated; meanwhile, the specific technological details like the determination of the optimal incubation pH value and the optimization of the pH adjusting mode of low pH incubation are optimized, and the efficiency and the effectiveness of the inactivation of common DNA viruses, RNA viruses, enveloped viruses and non-enveloped viruses are improved.

Description

technical field [0001] The invention belongs to the field of biotechnology. Specifically, the invention relates to a verification method for virus inactivation in low-pH incubation used in the production process of genetic engineering drugs / biological products. technical background [0002] With the development of biotechnology in the 21st century, the number of biological products produced by animal-derived tissues, cells, body fluids, and genetically engineered eukaryotic cells has gradually increased, and the number of people using them has continued to expand. In addition, the quality control of raw materials for animal-derived products has not attracted enough attention. The production process has not been strictly verified for virus removal / inactivation effect. Therefore, the risk of animal-derived viruses infecting humans is extremely high, and the problem of potential pathogenic infections has become increasingly prominent. Among them, due to the poor health and qua...

Claims

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Application Information

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IPC IPC(8): C12N7/06C12Q1/70C12Q1/6851C12Q1/18C12R1/93
CPCC12N7/00C12N2710/16763C12N2740/13063C12N2750/14363C12N2760/20263C12Q1/18C12Q1/6851C12Q1/701C12Q1/702C12Q1/705C12Q2531/113C12Q2563/107
Inventor 袁冰李姣丛娜刘传桂
Owner CANVEST WUHAN BIOTECH
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