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US 11,883,483 B2Granted
Method for preparing influenza working virus seed stock, method for preparing influenza vaccine using same seed stock, and virus seed stock prepared by same method
AssigneeSK Bioscience Co., Ltd.
InventorsYun Hee Kim, Yong Wook Park, Dong Soo Ham, Hwan Ui Jung, Hun Kim
Life of this patent
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Abstract
The present invention relates to a method of preparing an influenza working virus seed stock, a method of increasing infectivity of an influenza working virus seed stock, a method of preparing an influenza vaccine using the seed stock, an influenza vaccine prepared by the method, and an influenza working virus seed stock having increased infectivity.
Claims12
A method of preparing an influenza working virus seed stock, the method comprising:
- a) infecting a cell line adapted to serum-free culture and suspension culture with an influenza vaccine virus such that the infecting is carried out at a plurality of dilution factors, relative to a proportion of the influenza vaccine used to perform the infecting, to obtain a plurality of infected cell lines at different dilution factors;
- b) culturing the plurality of infected cell lines, to obtain a plurality of virus cultures at the different dilution factors;
- c) measuring cell viability of each of the plurality of virus cultures, and performing a hemagglutination assay (HA) on each of the plurality of virus cultures;
- d) selecting one of the plurality of virus cultures as a selected virus culture based on Condition 1 or Condition 2:
- Condition 1:
- if measured HA titers (except for HA titers of 0) of at least two of the plurality of virus cultures are different by a factor of 4 times or more, then the selected virus culture is a virus culture with a dilution factor having a highest measured HA titer, or
- Condition 2:
- if Condition 1 is not satisfied, then the selected virus culture is a virus culture with a dilution factor having a highest measured cell viability;
- and
- e) further passaging the selected virus culture in the same cell line at least once.
7 more elements
A method of increasing the infectivity of an influenza working virus seed stock, the method comprising:
- a) infecting a cell line adapted to serum-free culture and suspension culture with an influenza vaccine virus such that the infecting is carried out at a plurality of dilution factors, relative to a proportion of the influenza vaccine used to perform the infecting, to obtain a plurality of infected cell lines at different dilution factors;
- b) culturing the plurality of infected cell lines, to obtain a plurality of virus cultures at the different dilution factors;
- c) measuring cell viability of each of the plurality of virus cultures, and performing a hemagglutination assay (HA) on each of the plurality of virus cultures;
- d) selecting one of the plurality of virus cultures as a selected virus culture based on Condition 1 or Condition 2:
- Condition 1:
- if measured HA titers (except for HA titers of 0) of at least two of the plurality of virus cultures are different by a factor of 4 times or more, then the selected virus culture is a virus culture with a dilution factor having a highest measured HA titer, or
- Condition 2:
- if Condition 1 is not satisfied, then the selected virus culture is a virus culture with a dilution factor having a highest measured cell viability;
- and
- e) further passaging the selected virus culture in the same cell line to prepare an influenza working virus seed stock at least once.
7 more elements
Dependencies are inferred from the claim text.