Optimization of
Serum-Free Culture Conditions for HEK293T Cell-Based Production of Humanized Anti-CD147
Antibody
Thongheang K., Thongkum W., Yasamut U. and Tayapiwatana C.
Res. J. Biotech.; Vol. 21(1); 232-238;
doi: https://doi.org/10.25303/211rjbt2320238; (2026)
Abstract
The production of antibodies in mammalian cells is essential for therapeutic development,
but optimizing culture conditions remains a challenge for scalable production. To
optimize the culture conditions for antibody production in HEK293T cell, humanized
anti-CD147 antibody (HuM6-1B9) produced by HuM6-1B9-expressing HEK293T cells was
used as a model. The cells (3.0 × 105 cells/mL) were seeded into T25 cell culture
flasks. The performance of three serum-free media (SFM)—CDM4HEK293™, CHO-S-SFM II
and EX-CELL™ 293—under direct and sequential adaptation were evaluated. Cell viability
and cell number were examined using the trypan blue exclusion method and secreted
HuM6-1B9 in the culture supernatant was quantified using indirect ELISA.
The results showed that CHO-S-SFM II achieved the highest HuM6-1B9 yield, especially
with direct adaptation. It maintained high cell viability (97%) and enhanced per-cell
antibody production. A strong positive correlation (r = 0.893, p < 0.05) between
cell number and HuM6-1B9 concentration highlighted the importance of viable cell
density. In conclusion, direct adaptation in CHO-S-SFM II maximizes HuM6-1B9 yield
per cell, offering a foundation for optimizing recombinant antibody production in
HEK293T cells.
