US 9,809,800 B2
Method for producing parvovirus having high infectivity titer
Koichiro Yanagida, Tokyo (JP)
Assigned to ASAHI KASEI MEDICAL CO., LTD., Tokyo (JP)
Appl. No. 14/646,115
Filed by ASAHI KASEI MEDICAL CO., LTD., Tokyo (JP)
PCT Filed Sep. 5, 2013, PCT No. PCT/JP2013/073906
§ 371(c)(1), (2) Date May 20, 2015,
PCT Pub. No. WO2014/080676, PCT Pub. Date May 30, 2014.
Claims priority of application No. 2012-256801 (JP), filed on Nov. 22, 2012.
Prior Publication US 2015/0299668 A1, Oct. 22, 2015
Int. Cl. C12N 5/10 (2006.01); C12N 7/00 (2006.01)
CPC C12N 7/00 (2013.01) [C12N 2750/14351 (2013.01)] 15 Claims
 
1. A method for producing a parvovirus in a culture supernatant by culturing host cells and a seed virus of the parvovirus in a culture substrate, comprising:
(a) calculating: a doubling time of the host cells during the log growth phase in the culturing; and a cell density of the host cells confluently grown in the culturing;
(b) inoculating the seed virus of the parvovirus into the culture substrate comprising a medium and the host cells having a cell density of 1/500 to 1/20 of the cell density of the host cells confluently grown, calculated in (a), to give a multiplicity of infection (MOI) of 0.0001 to 0.1;
(c) culturing the culture comprising the medium and the host cells having a cell density of 1/500 to 1/20 of the cell density of the host cells confluently grown and the inoculated parvovirus obtained in (b) for a period of 5 to 11 times the doubling time calculated in (a); and
(d) recovering a culture supernatant comprising the parvovirus, obtained by the culturing (c).