Merten O W, Keller H, Cabanié L, Leno M, Hardefelt M
Institut Pasteur, Laboratoire de Technologie Cellulaire, Paris, France.
Cytotechnology. 1990 Jul;4(1):77-89. doi: 10.1007/BF00148813.
Pulse chase experiments of two mouse hybridoma lines were conducted in order to elucidate the kinetics of monoclonal antibody (mAb) production and secretion during different stages of batch cultures. The results indicate that a stock of cytoplasmic IgG exists in hybridoma cells and that the concentration of this stored IgG depends on the cell line used and the stage of the culture. This stored IgG can be released by dying cells, and a certain quantity of the secreted IgG is derived from this source. However, only between 0.3 and 9.3% of the released IgG of U0208 (average: 2.08%) and between 2.08 and 25.8% of the IgG, released from I.13.17 (average: 6.95%), were of storage origin, calculated on culture viability and intracellular IgG-stock. Comparing the accumulation of radio-labelled IgG (IgG*) in the supernatant with the reduction of cytoplasmic IgG* during the chase experiments, the percentages range between 14 and 50%, somewhat higher values probably caused by changes in the culture conditions. These changes led to a release of IgG during the chase experiments, which accounts for about 20-25% of the totally secreted IgG. It could be established that during the logarithmic growth phase of batch cultures a certain percentage of synthesized IgG was not released but stored within the cells: for U0208: 0.3-4.5%, for I.13.17: 1-7.6%. During the stationary and death phase, this percentage ranged between 1.5 and 20% for U0208 and between 0.5 and 8.1% for I.13.17. Finally, the chase experiments also revealed that the time of synthesis, assembly, and secretion of mAbs does not vary much during the different phases of batch cultures, and is within the range of 1.5 and 3 hrs.
为了阐明分批培养不同阶段单克隆抗体（mAb）产生和分泌的动力学，对两种小鼠杂交瘤细胞系进行了脉冲追踪实验。结果表明，杂交瘤细胞中存在细胞质IgG储备，且这种储存的IgG浓度取决于所用细胞系和培养阶段。这种储存的IgG可由死亡细胞释放，一定量的分泌型IgG即来源于此。然而，根据培养活力和细胞内IgG储备计算，U0208释放的IgG中只有0.3%至9.3%（平均为2.08%）以及I.13.17释放的IgG中有2.08%至25.8%（平均为6.95%）来自储存源。在追踪实验中，将上清液中放射性标记IgG（IgG*）的积累与细胞质IgG*的减少进行比较，其百分比在14%至50%之间，较高的值可能是由培养条件的变化引起的。这些变化导致在追踪实验期间IgG释放，约占总分泌IgG的20%至25%。可以确定，在分批培养的对数生长期，一定比例的合成IgG未释放而是储存在细胞内：U0208为0.3%至4.5%，I.13.17为1%至7.6%。在稳定期和死亡期，U0208的这一比例在1.5%至20%之间，I.13.17在0.5%至8.1%之间。最后，追踪实验还表明，单克隆抗体的合成、组装和分泌时间在分批培养的不同阶段变化不大，在1.5至3小时范围内。
