Abstract
Mammalian cell line stability is an important consideration when
establishing a biologics manufacturing process in the biopharmaceutical
and in vitro diagnostics (IVD) industries. Traditional CHO cell line
development methods use a random integration approach that requires
transfection, selection, optional amplification, screenings, and
single-cell cloning to select clones with acceptable productivity,
product quality, and genetic stability. Site-specific integration
reduces these disadvantages, and new technologies have been developed to
mitigate risks associated with genetic instability. In this study, we
applied the Leap-In® transposase-mediated expression system from ATUM to
generate stable CHOK1 pools for the production of four recombinant
antibody reagents for IVD immunoassays. CHO cell line stability is
defined by consistent antibody production over time. Three of the CHOK1
pools maintained productivity suitable for manufacturing, with high
antibody yields. The productivity of the remaining CHOK1 pool decreased
over time; however, derivative clones showed acceptable stability.
L-glutamine had variable effects on CHOK1 cell line or stable pool
stability and significantly affected antibody product titer. Compared
with traditional random integration methods, the ATUM Leap-In system can
reduce the time needed to develop new immunoassays by using semi-site
specific integration to generate high yield stable pools that meet
manufacturing stability requirements.