Preparation method of TGEV
A virus and microcarrier technology, applied in the field of cell engineering, can solve problems such as low titer of TGEV virus, and achieve the effects of uncomplicated cell harvesting process, low production cost and high cell yield
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Examples
Embodiment 1
[0031] Bioreactor: 14L bioreactor from Berenger, Germany.
[0032] Microcarrier: Cytodex-1 (purchased from GE).
[0033] Porcine transmissible gastroenteritis virus: TGEVHB08 strain.
[0034] Cell growth medium: MEM containing 8% newborn bovine serum by volume;
[0035] Virus maintenance solution: MEM containing 2% newborn bovine serum by volume;
[0036] Microcarrier suspension culture of ST cells: In a 14L bioreactor, add the treated microcarrier Cytodex-1 at a concentration of 2g / L, and inoculate ST cells with a cell inoculation concentration of 400,000 / mL. The optimal setting parameters of the reactor during cell culture: pH7.25, temperature 37°C, dissolved oxygen 55%, stirring speed 35rpm / min. After ST cells were inoculated, samples were taken at regular intervals, the state of the cells was observed under a microscope, and the cells were counted to determine the consumption of glucose. According to the situation of glucose consumption and cell growth and detachment, ...
Embodiment 2
[0040] Bioreactor: 14L bioreactor from Berenger, Germany.
[0041] Microcarrier: Cytodex-1 (purchased from GE).
[0042] Porcine transmissible gastroenteritis virus: TGEVHB08 strain.
[0043] Cell growth medium: MEM containing 10% newborn calf serum by volume;
[0044] Virus maintenance solution: MEM containing 3% newborn bovine serum by volume;
[0045] Microcarrier suspension culture of ST cells: In a 14L bioreactor, add the treated microcarrier Cytodex-1 at a concentration of 8g / L, and inoculate ST cells with a cell inoculation concentration of 1 million / mL. The optimal setting parameters of the reactor during cell culture: pH7.0, temperature 37°C, dissolved oxygen 55%, stirring speed 80rpm / min. After ST cells were inoculated, samples were taken at regular intervals, the state of the cells was observed under a microscope, and the cells were counted to determine the consumption of glucose. According to the glucose consumption and the growth and shedding of cells, the mediu...
Embodiment 3
[0049] A kind of TGEV virus preparation method, comprises the following steps:
[0050] 1) Get the microcarrier and place it in PBS buffer solution for soaking treatment, then sterilize, discard the PBS buffer solution, add it to the MEM cell growth solution containing 9% (w / w) serum, and obtain the mixed culture medium for subsequent use. The density of the microcarrier in the medium is 4g / L;
[0051] 2) Inoculate 0.6×10 6 Cells / mL ST cells, and then cultured at pH 6.6, temperature 35°C, dissolved oxygen 40%, stirring speed 35rpm;
[0052] 3) When the cell density reaches 5×10 6 Cells / mL, discard the cell growth solution in the mixed medium, add the virus maintenance solution, and then inoculate it with 1.5% porcine transmissible gastroenteritis virus, at a temperature of 36°C, dissolved oxygen 40%, and a stirring speed of 35rpm Under culture, wherein the virus maintenance solution is MEM cell growth solution containing 1% serum;
[0053] 4) When the virus is inoculated a...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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