Cell line development platform optimization for monoclonal antibody production in CHO-DG44

3^rd International conference on Bioprocess and Biosystems Engineering

September 14-15, 2015 Baltimore, USA

Elizabeth H Scheideman

NIH-National Institute of Allergy and Infectious Diseases, USA

Posters-Accepted Abstracts: J Bioproces Biotechniq

Abstract :

The Vaccine Production Program Laboratory at the National Institutes of Health translates research products from the Vaccine Research Center into material for proof-of-concept clinical trials. To meet production demands of >4 g/L for monoclonal antibodies, an efficient and reliable process for stable cell line generation is required. Evaluation of the early steps of the process used for VRC CHO-DG44 cells has been undertaken with a focus on transfection and MTX amplification. Transfection efficiencies and cell viabilities of >90% have been achieved through optimization of AmaxaTMNucleofectorTM and MaxCyte�® STXTM transfection and post-transfection recovery conditions. Additionally, replacing the ActiCHO production medium used for post-transfection culture with HyCloneTM CDM4CHOTM medium was found to increase MTX selection sensitivity and improve titers in shake flask batch cultures by 3- to 6-fold. Finally, a monoclonal antibody-producing clone was subjected to varying concentrations of MTX (from 30-300 nM). Cells from wells with the highest titer after 1-2 weeks in 24-well plates were pooled and expanded. In un-optimized shake flask fed-batch runs, product titers for cells grown in 100 or 300 nM MTX were increased by approximately 1.5- to 2-fold compared to those in 30 nM MTX (from 0.7 g/L to 1.2-1.4 g/L). Together, these modifications to the existing process are critical to ensure that subsequent clone selection will provide stable, high-producing cell lines.

Biography :

Email: elizabeth.scheideman@nih.gov