US 11,781,113 B2
End-to-end cell therapy automation
Yaling Shi, Walkersville, MD (US); Erika McAfee, Walkersville, MD (US); Samatha Bandapalle, Walkersville, MD (US); Ann Siehoff, Cologne (DE); Timo Gleissner, Cologne (DE); Joseph O'Connor, Walkersville, MD (US); Eytan Abraham, Walkersville, MD (US); Kelly Purpura, Kingston (CA); Nuala Trainor, Kingston (CA); and Timothy Smith, Kingston (CA)
Assigned to LONZA WALKERSVILLE, INC., Walkersville, MD (US); LONZA COLOGNE GMBH, Cologne (DE); and OCTANE BIOTECH INC., Kingston (CA)
Filed by LONZA WALKERSVILLE, INC., Walkersville, MD (US); LONZA COLOGNE GMBH, Cologne (DE); and OCTANE BIOTECH INC., Kingston (CA)
Filed on May 24, 2022, as Appl. No. 17/752,021.
Application 17/752,021 is a continuation of application No. 16/119,618, filed on Aug. 31, 2018, granted, now 11,447,745.
Claims priority of provisional application 62/670,391, filed on May 11, 2018.
Claims priority of provisional application 62/553,214, filed on Sep. 1, 2017.
Prior Publication US 2022/0282214 A1, Sep. 8, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/0783 (2010.01); C12M 3/00 (2006.01); C12M 1/00 (2006.01); C12M 1/34 (2006.01); C07K 14/725 (2006.01); C07K 14/705 (2006.01); C12N 5/00 (2006.01); C12N 5/16 (2006.01); C12N 15/86 (2006.01)
CPC C12N 5/0636 (2013.01) [C07K 14/7051 (2013.01); C07K 14/70521 (2013.01); C12M 23/42 (2013.01); C12M 29/20 (2013.01); C12M 41/00 (2013.01); C12N 5/0018 (2013.01); C12N 5/163 (2013.01); C12N 15/86 (2013.01); C12N 2510/00 (2013.01); C12N 2740/10041 (2013.01); C12N 2740/15041 (2013.01)] 17 Claims
 
1. A method for automated production of a genetically modified T cell culture, the method comprising:
a) mixing a T cell culture with a magnetic selection reagent;
b) exposing the T cell culture to magnetic separation;
c) activating the T cell culture with an activation reagent selected from an antibody and a dendritic cell to produce an activated T cell culture;
d) transducing within a cell culture chamber the activated T cell culture with a viral vector encoding an ectodomain, a transmembrane domain and an endodomain, to produce a transduced T cell culture;
e) expanding within the cell culture chamber the transduced T cell culture;
f) centrifuging the expanded T cell culture; and
g) harvesting the T cell culture to produce a genetically modified T cell culture,
wherein (a) through (f) are performed within a fully enclosed, automated cell engineering system, and the cell culture chamber has a fixed area; and
wherein expansion of the transduced T cell culture in (c) produces at least 20% more genetically modified T cells than expansion utilizing manual cell culture with a flexible, gas permeable bag and transduction efficiency of the method is at least 20% higher than transduction efficiency utilizing manual cell culture with the flexible, gas permeable bag.