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\begin{document}

\title{Optimization of a transient antibody expression platform towards high
titer and efficiency}



\author[1]{Elizabeth Greene}%
\author[1]{Ninkka Tamot}%
\author[1]{Daniela Cazacu}%
\author[1]{Douglas Gebhard}%
\author[1]{Steven Castellano}%
\author[1]{Anthony Kronkaitis}%
\author[1]{Jon Reed}%
\author[1]{Paul Mason}%
\author[1]{Lore Florin}%
\author[1]{Nicholas Rossi}%
\author[1]{Anthony Lauer}%
\author[1]{Laura Juckem}%
\author[1]{Andrew Nixon}%
\author[2]{Till Wenger}%
\author[3]{Saurabh Sen}%
\affil[1]{Affiliation not available}%
\affil[2]{Boehringer Ingelheim}%
\affil[3]{Boehringer Ingelheim Corp USA}%


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  \date{June 14, 2024}


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\begin{abstract}
Transient gene expression (TGE) using mammalian cells is an extensively
used technology for production of antibodies and recombinant proteins
and has been widely adapted by both academic and industrial labs.
Chinese Hamster Ovary (CHO) cells have become one of the major work
horses for TGE of recombinant antibodies due to their attractive
features: post-translational modifications, adaptation to high cell
densities, and use of serum-free media. In this study, we describe the
optimization of parameters for TGE for antibodies from CHO cells.
Through a matrix evaluation of multiple factors including inoculum,
transfection conditions, amount and type of DNA used and
post-transfection culture conditions, we arrived at an optimized process
with higher titer and reduced costs and time, thus increasing the
overall efficiency of early antibody material supply. We investigated
the amount of coding DNA and the influence of size of the transfection
complex on the in vitro efficiency of the transfection. Generation of
the transfection complex in serum-free medium leads to the prompt
formation of an optimal-sized polyplex, and is independent of the
relative amount of coding DNA used for a successful transfection
outcome.%
\end{abstract}%



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