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Method for removing residual host DNA and host protein in encephalitis B vaccine product

A technology for host proteins and vaccines, applied in the field of virus vaccine product purification, can solve problems such as adverse reactions, causing cancer, side effects, etc., and achieve the effect of improving product quality standards and breaking through quality bottlenecks

Pending Publication Date: 2022-04-26
LIAONING CHENGDA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The production of JE vaccine is to inoculate the virus on suitable animal cells, release the virus into the culture medium after propagation, and at the same time, some residual host cells or their fragments will also enter the culture medium, after clarification, super After concentration by filtration and gel filtration chromatography, free host DNA and host protein and host DNA and host protein combined with antigenic protein will be produced in the extracted JE vaccine stock solution; among them, ultrafiltration concentration and gel filtration chromatography process A certain proportion of free DNA and host protein will be removed, but there will still be residual DNA and host protein, especially the DNA and host protein combined with virus or antigenic protein will remain in the stock solution. If the removal is not complete, this part DNA and host protein will be injected into the human body together with the vaccine, which may cause adverse reactions in the human body, and may lead to cancer in severe cases
[0003] At present, JE vaccines are generally extracted by traditional processes such as concentration and gel filtration chromatography. The resulting JE vaccine has the following defects: the residual host DNA content after the virus vaccine is extracted is too high; the virus vaccine still contains too high host protein, and the removal rate of miscellaneous proteins is not high, resulting in relatively large side effects after clinical use of the vaccine
The above defects have affected the quality of virus vaccine products and restricted the production of vaccine products
[0004] Multimodal chromatography packing material Capto MMC is a complex ligand cation exchange chromatography medium, which is suitable for fast, efficient and cost-effective protein purification process. However, there is no use of multimodal chromatography packing material Capto MMC in related technologies. Related reports on purification methods of vaccine products

Method used

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  • Method for removing residual host DNA and host protein in encephalitis B vaccine product
  • Method for removing residual host DNA and host protein in encephalitis B vaccine product
  • Method for removing residual host DNA and host protein in encephalitis B vaccine product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Use AKTA avant 150 automatic chromatography equipment to pass 200mL JE vaccine concentrated inactivation solution (code A1) through the XK16 chromatography column for multi-mode Capto MMC chromatography, the ultraviolet detection wavelength is 280nm, the flow rate is 1.49cm / min, and the mobile phase 0.01mol / L PBS buffer solution (pH7.5, NaCl 0.15mol / L), collect the first absorption peak, that is, the one-step purification solution of Japanese encephalitis virus protein (number is A2, see figure 1). Then one-step purified solution was subjected to Sepharose 6 Fast FLow gel filtration chromatography, the ultraviolet detection wavelength was 280nm, the flow velocity was 25cm / h, and the mobile phase was 0.01mol / L PBS buffer solution (pH7.5, NaCl0.15mol / L), and collected The first absorption peak is the final purified solution of JEV protein (numbered A3, see figure 2 ).

[0032] A1 and A3 were tested for antigen content, DNA residue, and host protein residue, respectivel...

Embodiment 2

[0038] Use AKTA avant 150 automatic chromatography equipment to pass 320mL JE vaccine concentrated inactivation solution (code B1) through the XK50 chromatography column for Sepharose 6 Fast FLow gel filtration chromatography, the ultraviolet detection wavelength is 280nm, and the flow rate is 25cm / h. The mobile phase is 0.01mol / L PBS buffer (pH7.5, NaCl 0.15mol / L), and the first absorption peak is collected, which is the one-step purification solution of Japanese encephalitis virus protein (numbering is B2, see image 3 ). Then the one-step purified solution (100mL) was subjected to multi-mode CaptoMMC chromatography, the ultraviolet detection wavelength was 280nm, the flow rate was 1.49cm / min, and the mobile phase was 0.01mol / L PBS buffer solution (pH7.5, NaCl 0.15mol / L), and collected The first absorption peak is the final purified solution of JEV protein (numbered as B3, see Figure 4 ).

[0039] B1 and B3 were tested for antigen content, DNA residue, and host protein re...

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Abstract

The invention relates to the field of purification of viral vaccine products, and particularly discloses a method for removing residual host DNA and host protein in an encephalitis B vaccine product. A multi-mode medium Capto MMC is combined with a gel filtration chromatography medium to purify the encephalitis B vaccine concentrated inactivation solution, so that the purpose of removing host DNA and host protein is achieved. The method has the advantages that the multi-mode medium Capto MMC and the gel filtration chromatography medium are combined for use, the concentrated inactivated liquid of the encephalitis B vaccine is purified, and the purpose of removing host DNA and host protein is achieved, so that the residual quantity of DNA and the residual quantity of host protein of the finished encephalitis B vaccine are further reduced, and the product quality of the encephalitis B vaccine is improved.

Description

technical field [0001] This application relates to the field of purification of virus vaccine products, more specifically, it relates to a method for removing residual host DNA and host protein from Japanese encephalitis vaccine products. Background technique [0002] The production of JE vaccine is to inoculate the virus on suitable animal cells, release the virus into the culture medium after propagation, and at the same time, some residual host cells or their fragments will also enter the culture medium, after clarification, super After concentration by filtration and gel filtration chromatography, free host DNA and host protein and host DNA and host protein combined with antigenic protein will be produced in the extracted JE vaccine stock solution; among them, ultrafiltration concentration and gel filtration chromatography process A certain proportion of free DNA and host protein will be removed, but there will still be residual DNA and host protein, especially the DNA a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/02
CPCC12N7/00C07K1/165C07K1/16C12N2770/24151C12N2770/24134Y02A50/30
Inventor 李庆岸杨兵于海曹鹏康文哲由迪
Owner LIAONING CHENGDA BIOTECH
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