Impact of N-1 phase media optimization as a bioprocess intensification strategy on fed-batch production.

Authors

Mustafa Doğukan Metiner, Elif Damla Arisan, Abdullah Uslu, Işıl Kurnaz

Published in

Bioprocess and biosystems engineering. Jun 25, 2026. Epub Jun 25, 2026.

Abstract

In the biopharmaceutical industry, improving process efficiency and reducing manufacturing costs remain major priorities. Conventional fed-batch processes are often limited by low initial cell densities, resulting in extended culture duration and reduced productivity. However, the combined effect of N-1 media optimization and high-cell-density (HCD) inoculation remains insufficiently characterized. This study systematically assessed the impacts of N-1 media optimization and initial seeding density utilizing a CHO-ZN cell line. During the N-1 phase, a media exchange strategy increased the cell density from approximately 3 × 10⁶ cells/mL to 15-22 × 10⁶ cells/mL, enabling direct production inoculation at 8 × 10⁶ cells/mL without an additional seed expansion step. Compared with low-cell-density (LCD; 1.2 × 10⁶ cells/mL) cultures, HCD cultures reduced production duration by approximately 36% and increased product concentration by about 89% (~ 3500 mg/L vs. ~1850 mg/L). Average interval-based specific productivity (qP), increased from approximately 17 to 31 pg/cell/day, suggesting that the gains in productivity were not solely attributable to increased biomass. Importantly, when the production-phase medium was standardized, differences in titer concentration, metabolite profiles, viability, and interval-based qP persisted among cells conditioned in different N-1 media. This indicates that N-1 medium composition influenced subsequent fed-batch behavior not only by affecting inoculum biomass, but also by modulating the physiological state of the cells prior to production inoculation. Overall, these findings emphasize the role of N-1 medium-dependent physiological conditioning in determining productivity, metabolic behavior, and product quality attributes under intensified fed-batch conditions.

PMID:
42347961
Bibliographic data and abstract were imported from PubMed on 25 Jun 2026.

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