US 12,264,324 B2
Viral vector assay and vector
Nigel Parker, Chinnor (GB); Hanna P. Lesch, Kuopio (FI); Jenni Mykkanen, Kuopio (FI); Sara Paulo, Kuopio (FI); Minna Hassinen, Kuopio (FI); and Robert Shaw, Kuopio (FI)
Assigned to Trizell Ltd., Chinnor (GB)
Filed by Trizell Ltd., Chinnor (GB)
Filed on Jul. 28, 2021, as Appl. No. 17/386,644.
Application 17/386,644 is a continuation of application No. 15/754,598, granted, now 11,104,915, previously published as PCT/US2016/050959, filed on Sep. 9, 2016.
Claims priority of provisional application 62/218,810, filed on Sep. 15, 2015.
Prior Publication US 2022/0081696 A1, Mar. 17, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/86 (2006.01); G01N 33/53 (2006.01); G01N 33/569 (2006.01)
CPC C12N 15/86 (2013.01) [G01N 33/56983 (2013.01); C12N 2710/10343 (2013.01); C12N 2710/10351 (2013.01)] 17 Claims
 
1. A method of assaying a batch of recombinant viral vectors to determine if the recombinant viral vectors can be packaged and released, wherein the recombinant viral vectors have a viral transgene that expresses a protein that has biological activity, said method comprising determining a level of infectivity of a sample of the recombinant viral vectors comprising:
a. infecting cultured cells with the sample of the recombinant viral vectors, wherein the cultured cells are infected with more than one concentration of the recombinant viral vector;
b. measuring by flow cytometry a percentage of infected cells that comprise the expressed recombinant viral vector protein;
c. using the percentage of infected cells that comprise the expressed recombinant viral vector protein in a Slope Ratio method to generate a linear response curve; and
d. comparing the linear response curve to a reference standard to determine level of infectivity,
wherein if the level of infectivity is acceptable, then the batch of recombinant viral vectors is packaged and released.