Zijlstra G M, Michielsen M J, de Gooijer C D, van der Pol L A, Tramper J
Bio-Intermediair Europe BV, Groningen, The Netherlands.
Biotechnol Prog. 1996 May-Jun;12(3):363-70. doi: 10.1021/bp960017e.
The partitioning of mouse/mouse hybridoma cell line BIF6A7, mouse/rat hybridoma PFU-83, and CHO DUKX B11-derived cell line BIC-2 in aqueous two-phase systems (ATPSs) of poly(ethylene glycol) (PEG) and dextran was studied. The partitioning of BIF6A7 was investigated systematically by using a statistical experimental design. The aims were to identify the key factors governing cell partitioning and to select ATPSs with suitable cell partitioning for extractive bioconversions with animal cells. The influence of five factors, i.e., the poly(ethylene glycol) molecular weight (PEG MW), dextran molecular weight (Dx MW), tie-line length (TLL), pH, and the ratio of potassium phosphate to potassium chloride, defined as the fraction KPi/(KPi + KCl), on BIF6A7 cell partitioning was characterized by using a full factorial experimental design. The cell partitioning ranged from complete partitioning into the interface to an almost complete partitioning to the lower phase. In all cases less than 1% of the cells partitioned to the top phase. The potassium phosphate fraction had the largest effect on cell partitioning. Low potassium phosphate fractions increased the proportion of cells in the lower phase. To a lesser extent the other factors also played a role in the cell partitioning. The best partitioning for the BIF6A7 cell line was obtained in ATPSs with PEG MW = 35,000, Dx MW = 40,000, TLL = 0.10 g/g, pH 7.4, and KPi/(KPi + KCl) = 0.1, where 93% of the cells were present in the lower phase. The previously reported partitioning of BIF6A7 cells in ATPS culture medium, corresponded well with the current findings. The partitioning of mouse/rat hybridoma cell line PFU-83 and CHO cell line BIC-2 was studied in an ATPS culture medium with PEG 35,000, dextran 40,000, TLL = 0.12 g/g, and hybridoma culture medium. Both cell lines partitioned almost completely into the lower phase. Moreover, the PFU-83 cell line was able to grow in the ATPS hybridoma culture medium. This ATPS hybridoma culture medium therefore seems to be suitable for extractive bioconversions with a wide range of hybridoma cell lines, provided that their product can be partitioned into the upper PEG-rich phase.
研究了小鼠/小鼠杂交瘤细胞系BIF6A7、小鼠/大鼠杂交瘤PFU - 83以及CHO DUKX B11衍生细胞系BIC - 2在聚乙二醇（PEG）和葡聚糖的双水相体系（ATPSs）中的分配情况。通过统计实验设计系统地研究了BIF6A7的分配情况。目的是确定控制细胞分配的关键因素，并选择具有合适细胞分配的双水相体系用于动物细胞的萃取生物转化。采用全因子实验设计，表征了聚乙二醇分子量（PEG MW）、葡聚糖分子量（Dx MW）、系线长度（TLL）、pH以及磷酸钾与氯化钾的比例（定义为KPi/(KPi + KCl)）这五个因素对BIF6A7细胞分配的影响。细胞分配范围从完全分配到界面到几乎完全分配到下相。在所有情况下，不到1%的细胞分配到上相。磷酸钾比例对细胞分配的影响最大。低磷酸钾比例增加了下相中细胞的比例。在较小程度上，其他因素也在细胞分配中起作用。对于BIF6A7细胞系，在PEG MW = 35000、Dx MW = 40000、TLL = 0.10 g/g、pH 7.4以及KPi/(KPi + KCl) = 0.1的双水相体系中获得了最佳分配，其中93%的细胞存在于下相。先前报道的BIF6A7细胞在双水相培养基中的分配情况与当前研究结果吻合良好。在含有PEG 35000、葡聚糖40000、TLL = 0.12 g/g的双水相培养基和杂交瘤培养基中研究了小鼠/大鼠杂交瘤细胞系PFU - 83和CHO细胞系BIC - 2的分配情况。两个细胞系几乎都完全分配到下相。此外，PFU - 83细胞系能够在双水相杂交瘤培养基中生长。因此，这种双水相杂交瘤培养基似乎适用于多种杂交瘤细胞系的萃取生物转化，前提是它们的产物能够分配到富含PEG的上相。
