US 12,487,241 B2
Rapid verification of virus particle production for a personalized vaccine
Adrian E. Rice, Culver City, CA (US); Kayvan Niazi, Culver City, CA (US); and Frank R. Jones, Culver City, CA (US)
Assigned to NantCell, Inc., Culver City, CA (US); and NantBio, Inc., Culver City, CA (US)
Filed by NANTCELL, INC., Culver City, CA (US); and NANTBIO, INC., Culver City, CA (US)
Filed on Dec. 16, 2022, as Appl. No. 18/067,508.
Application 18/067,508 is a continuation of application No. 16/964,325, granted, now 11,573,230, previously published as PCT/US2019/015126, filed on Jan. 25, 2019.
Claims priority of provisional application 62/622,758, filed on Jan. 26, 2018.
Prior Publication US 2023/0213518 A1, Jul. 6, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 19/34 (2006.01); A61K 39/00 (2006.01); C12N 15/86 (2006.01); G01N 33/569 (2006.01)
CPC G01N 33/56983 (2013.01) [A61K 39/00 (2013.01); C12N 15/86 (2013.01); C12N 2710/10334 (2013.01); C12N 2710/10343 (2013.01); C12N 2710/10351 (2013.01)] 13 Claims
OG exemplary drawing
 
1. A method of producing a neoantigen vector for use in generating a neoantigen vaccine for a subject in need thereof, the method comprising:
transfecting a plasmid into adenoviral production cells, wherein the plasmid comprises (1) a polynucleotide encoding adenoviral proteins and (2) a polynucleotide insert encoding at least one tumor-derived neoantigen, thereby producing and propagating adenoviral vectors in the adenoviral production cells;
lysing the adenoviral production cells, wherein the lysate comprises the adenoviral vectors and adenovirus-associated hexon protein;
loading the lysate onto an optical biosensor comprising a glass surface and an anti-hexon monoclonal antibody;
measuring biomass accumulation on the glass surface by bio-layer interferometry (BLI), wherein the accumulation correlates with adenovirus-producing hexon positive production cells;
quantifying hexon positive production cells, wherein quantification of the purified lysate by BLI occurs within 5-30 minutes, and is correlated with infectivity of the adenoviral vector;
and thereby confirming production of infective adenoviral vectors for further propagation in a suspension cell line.