Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A kind of concentration method of virus liquid

A technology of virus liquid and virus, which is applied in the concentration of virus liquid and the field of virus culture, can solve the problems of inability to obtain virus, discomfort of virus particles, and harmfulness of infected cells. foreground effect

Active Publication Date: 2022-01-11
苏州良辰生物医药科技有限公司
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is required that the titer of the indicator virus must reach 106TCID50 or more, but different viruses have different proliferation effects on cells, and conventional proliferation and culture cannot obtain high-titer viruses, so necessary concentration methods must be adopted to increase the virus titer
[0004] As a method of increasing the concentration of viruses in a solution and concentrating viruses, there are physical methods such as ultracentrifugation, but this method requires expensive equipment, a long separation time, and requires a lot of labor in the operation
There are also chemical methods, such as the method of concentrating the virus by precipitating the virus with ammonium sulfate, polyethylene glycol, etc., but in these methods, because the reagents used or other components for precipitation are harmful to the infected cells, there are obstacles that hinder subsequent operations. Hidden dangers and tedious work such as purification must be carried out after virus precipitation
In addition, there is a method of using antibodies to concentrate the virion itself, but this method inhibits the function of the protein on the surface of the virus, which may affect the infectivity of the virus. For example, when separating the virion from the antibody, strong acidic conditions must be used (pH2~3) for elution, which has the potential to cause discomfort to virus particles

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of concentration method of virus liquid
  • A kind of concentration method of virus liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Cell subculture: Take out 1 tube of frozen ST cells from liquid nitrogen, thaw quickly in a 37°C water bath, use a sterile pipette to draw 1.0ml of cell suspension into a 15ml sterile centrifuge tube, and add drop by drop MEM medium containing 10wt% FBS to 5-15mL, pipet several times, mix evenly, centrifuge immediately (3000rpm, 5min), and discard the supernatant. Then add 10-20mL of 10wt% FBS MEM medium, pipette several times, mix well, transfer to T25 culture bottle with 10wt% FBS MEM medium, place at 37°C, 5% CO 2 Cultivate in an incubator for 48h to 72h, and when the cells cover a monolayer, set aside.

[0023] 2. Subculture of the virus: Take out 1 tube of frozen PPV virus from the -70°C refrigerator (working virus bank), redissolve in a 37°C water bath, dilute 10 times with MEM medium without FBS, and then inoculate 1mL in In the cell bottle that has been covered with a monolayer of cells, store at 37°C, 5% CO 2 The incubator was adsorbed for 1 hour, and the ...

Embodiment 2

[0029] 1. Cell subculture: Take out 1 tube of frozen LLC-MK2 cells from liquid nitrogen, thaw quickly in a 37°C water bath, use a sterile pipette to draw 1.0ml of cell suspension into a 15ml sterile centrifuge tube, drop by drop Add dropwise DMEM medium containing 10wt% FBS to 5-15mL, pipette several times, mix well, centrifuge immediately (3000rpm, 5min), and discard the supernatant. Then add 10-20 mL of DMEM medium with 10wt% FBS, pipette several times, mix well, transfer to T25 culture bottle with DMEM medium with 10wt% FBS, place at 37°C, 5% CO 2 Cultivate in an incubator for 24h to 48h, and when the cells cover a single layer, set aside.

[0030] 2. Subculture of the virus: take out a tube of frozen ReoV-3 virus from the -70°C refrigerator (working virus bank), redissolve it in a 37°C water bath, dilute it 10 times with MEM medium without FBS, and then pipette 1mL Inoculate in a cell bottle that has been filled with a monolayer of cells, store at 37°C, 5% CO 2 The incub...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention relates to a method for concentrating virus liquid, which comprises placing a biofilm in a vessel, and then adding 18-25ml of virus liquid to the vessel according to the weight of 1 g of the biofilm, and adding the virus liquid to the vessel, After adsorption at 2-8°C for 1-3 hours, freeze-drying was performed, and then reconstituted in MEM medium containing 1-3wt% fetal bovine serum, the biofilm was cut into pieces and homogenized, and then the biofilm was removed by centrifugation Fragments, the supernatant was filtered to obtain the concentrated virus liquid. The concentration method of the present invention is simple and feasible, and the concentrated virus liquid maintains the virus infectivity while improving the virus titer (not less than 10 7.0 TCID 50 / 0.1ml), and the virus liquid in the isolated state can be obtained, which has wide application prospects in the verification of virus removal / inactivation.

Description

technical field [0001] The invention belongs to virus culture technology in the field of microbe application, and in particular relates to a method for concentrating virus liquid. Background technique [0002] Biopharmaceutical products such as monoclonal antibodies, recombinant proteins, vaccines, blood derivatives, and animal products carry the risk of transmitting infectious viruses because the source material may itself be contaminated with viruses or virus-like particles. Furthermore, the production process of biopharmaceutical products is susceptible to viral contamination from external sources. Manufacturers of biopharmaceutical products therefore need to include adequate virus removal steps in their manufacturing processes to ensure that their products are free of contaminating viruses. [0003] The most commonly used viral assays to quantify viral contaminants in viral clearance studies are the tissue culture limiting dose 50% (TCID50), plaque forming unit (PFU) an...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/02
CPCC12N7/00
Inventor 吴娟缪汝娉鲍大为
Owner 苏州良辰生物医药科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com