Evaluation of anchorage-dependent cell propagation systems for production of human acetylcholinesterase by recombinant 293 cells.
Production of recombinant human acetylcholinesterase (AChE) by a high producer human embryonic kidney cell line (293) was evaluated by three main cell propagation systems; surface propagator, fixed-bed reactor and stirred microcarrier cultures. The recombinant cell line expresses AChE levels as high as 10-20 mg/l/day. System productivities in either the surface propagator (multitray system), or in the fixed-bed reactor (polyurethane macroporous sponges) were 4-8 mg AChE/l/day during a production period of 8 days. Similar productive rates, yet longer production periods (up to 22 days), were obtained in microcarrier (MC) cultures using either polystyrene beads (Biosilon); collagen-coated dextran beads (Cytodex-3); or gelatin macroporous beads (Cultispher-G). Best results were obtained in an aggregate culture using cellulose beads charged with diethylaminoethyl (DEAE) groups, (Servacel), as carriers. In this culture, a system productivity of 6-10 mg/l/day was maintained for 28 days.
Lazar
A
A
Department of Biotechnology, Israel Institute for Biological Research, Ness-Ziona.
Reuveny
S
S
Kronman
C
C
Velan
B
B
Shafferman
A
A
D000110:Acetylcholinesterase / Q000096:biosynthesis*; D001709:Biotechnology / Q000379:methods*; D002455:Cell Division; D002460:Cell Line; D005069:Evaluation Studies as Topic; D006801:Humans; D007668:Kidney / Q000166:cytology; D011994:Recombinant Proteins / Q000096:biosynthesis
D011994:Recombinant Proteins; D000110:Acetylcholinesterase