US 11,939,585 B2
Multi-pulse transfection methods and cells
Marcos Sixto, Culver City, CA (US); Kayvan Niazi, Culver City, CA (US); Ting Wu, Culver City, CA (US); and Chihwei Chang, Culver City, CA (US)
Assigned to NantBio, Inc., Culver City, CA (US)
Filed by NantBio, Inc., Culver City, CA (US)
Filed on Jun. 17, 2020, as Appl. No. 16/903,971.
Application 16/903,971 is a continuation of application No. 15/727,150, filed on Oct. 6, 2017, granted, now 10,724,043.
Claims priority of provisional application 62/404,993, filed on Oct. 6, 2016.
Prior Publication US 2020/0318126 A1, Oct. 8, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/65 (2006.01); C12N 5/10 (2006.01); C12N 15/82 (2006.01); C12N 15/85 (2006.01); C12N 15/87 (2006.01); A61K 48/00 (2006.01)
CPC C12N 15/8206 (2013.01) [C12N 5/10 (2013.01); C12N 15/85 (2013.01); C12N 15/87 (2013.01); A61K 48/00 (2013.01)] 16 Claims
 
1. A method of transfecting mammalian cells with a RNA, comprising:
disposing the mammalian cells with the RNA in a transfection medium, wherein the mammalian cells are immune competent cells
applying 2, 3 or 4 pulses to the mammalian cells at a field strength of 1,200 V/cm and a time constant from about 0.5 msec to about 5 msec to produce transfected mammalian cells,
wherein the transfected mammalian cells have a viability of at least 70%.