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Membrane anchored protein production from spheroid, porous, and solid microcarrier chinese hamster ovary cell cultures
Author(s) -
Kennard M. L.,
Piret J. M.
Publication year - 1995
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260470507
Subject(s) - microcarrier , chinese hamster ovary cell , cell culture , cell , microbiology and biotechnology , recombinant dna , chemistry , biology , biochemistry , gene , genetics
The influence of the microcarrier type on the performance of a controlled release process used to produce a recombinant glycosyl‐phosphatidylinositol anchored protein was investigated. Chinese hamster ovary (CHO) cells expressing the human melanoma tumor antigen (p97) were cultured in 10% serum on Cultispher‐GH porous microcarriers and then, for protein production, maintained in 2% serum. Cells were harvested every 48 h and p97 was recovered at 90 μg/mL and 40% purity. Harvested p97 concentrations were increased by harvestingfrom spheroid (241 μg/mL) and smaller porous microcarrier, Cultispher‐G (167 μg/mL) cultures. The low total cell specific p97 production of cells cultured on Cultispher‐GH was due to necrosis of cells within the beads, decreased p97 expression of the immobilized cells, dilution by the liquid (up to 40% volume) associated with settled beads, and incomplete recovery of p97 from within the beads. Cells cultured on solid microcarriers, Cytodex‐1, had the highest cell viability and cell specific p97 production, It is recommended that a two‐stage cyclic harvesting process of cells cultured on small Cultispher‐G or on Cytodex‐1 beads would minimize protein loss and maximize cell specific protein recovery. © 1995 John Wiley & Sons Inc.