Optimization of a transient antibody expression platform towards high titer and efficiency

Autor: Anthony Kronkaitis, Steven Castellano, Nicholas Rossi, Jon Reed, Anthony Lauer, Akshita Datar, Daniela Cazacu, Paul Mawson, Elizabeth Greene, Saurabh Sen, Laura Juckem, Andrew E. Nixon, Till Wenger, Ninkka Tamot, Lore Florin, Douglas Gebhard
Rok vydání: 2020
Předmět:
Zdroj: Biotechnology journalREFERENCES. 16(4)
ISSN: 1860-7314
Popis: Transient gene expression (TGE) using mammalian cells is an extensively used technology for the production of antibodies and recombinant proteins and has been widely adopted by both academic and industrial labs. Chinese Hamster Ovary (CHO) cells have become one of the major workhorses 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 uniquely optimized process with higher titer and reduced costs and time, thus increasing the overall efficiency of early antibody material supply. We further investigated the amount of coding DNA used in TGE and the influence of kinetics and size of the transfection complex on the in vitro efficiency of the transfection. We present here the first report of an optimized TGE platform using Filler DNA in an early drug discovery setting for the screening and production of therapeutic mAbs.
Databáze: OpenAIRE
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