Mar 18, 2005

Cytochalasin D can improve heterologous protein productivity in adherent Chinese hamster ovary cells

Biotechnology and Bioengineering
Eric J Hayduk, Kelvin H Lee


We generated a series of adherent gene-amplified CHO clones expressing human secreted alkaline phosphatase (SEAP) as a model for heterologous protein production. Clones demonstrate a 26- to 52-fold increase in productivity compared to controls after dhfr/methotrexate-mediated gene amplification and clone selection. SEAP is stably expressed in these clones over at least a 6-week period without significant productivity loss. Two-dimensional protein electrophoresis identified 21 proteins that exhibited altered expression in clones of increasing SEAP productivity. Based on MALDI TOF/TOF mass spectrometry of relevant protein spots, changes in translation, energy pathways, chaperones, regulatory proteins, and cytoskeletal proteins were observed, including a 4-fold expression increase in actin capping protein. We hypothesized that an alteration of the actin cytoskeleton using cytochalasin D as a mimic for actin-capping protein could have a beneficial effect on heterologous protein secretion. Treatment with 0.5 mug/mL cytochalasin D increased SEAP productivity 2- to 3-fold compared to an amplified control which resulted in an increase in productivity from 52- to 150-fold compared to a nonamplified parent.

Mentioned in this Paper

Ovarian Diseases
Chinese Hamster
Gene Amplification
Both Ovaries
Proteins, Recombinant DNA
Cell Culture Techniques
Chinese Hamster Ovary Cell
Actin Capping Proteins

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