Lazar A, Reuveny S, Kronman C, Velan B, Shafferman A
Department of Biotechnology, Israel Institute for Biological Research, Ness-Ziona.
Cytotechnology. 1993;13(2):115-23. doi: 10.1007/BF00749938.
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.
通过三种主要的细胞增殖系统,即表面增殖器、固定床反应器和搅拌微载体培养,对高产人胚胎肾细胞系(293)生产重组人乙酰胆碱酯酶(AChE)进行了评估。该重组细胞系表达的AChE水平高达10 - 20毫克/升/天。在8天的生产期内,表面增殖器(多盘系统)或固定床反应器(聚氨酯大孔海绵)中的系统生产率为4 - 8毫克AChE/升/天。使用聚苯乙烯珠(Biosilon)、胶原包被的葡聚糖珠(Cytodex - 3)或明胶大孔珠(Cultispher - G)的微载体(MC)培养获得了相似的生产率,但生产周期更长(长达22天)。使用充有二乙氨基乙基(DEAE)基团的纤维素珠(Servacel)作为载体的聚集体培养获得了最佳结果。在这种培养中,系统生产率为6 - 10毫克/升/天,持续了28天。