Hayduk EJ, Lee KH.Cytochalasin D can improve heterologous
protein productivity in adherent Chinese hamster ovary cells.
Biotechnol Bioeng. 2005 May 5;90(3):354-64
School of Chemical and Biomolecular Engineering, Cornell
University, Ithaca, New York 14853, USA.
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. Copyright 2005 Wiley Periodicals, Inc.
PMID: 15772946 |
