Laboratoire des Sciences du Génie Chimique, CNRS-INPL, BP 451, F-54001, Nancy Cedex, France.
Cytotechnology. 2000 Jan;32(1):17-29. doi: 10.1023/A:1008069523163.
Experiments, earlier performed in our laboratory, showedthe stimulating effect of butyric acid on monoclonalantibody production by hybridoma cells. Itssimulaneous inhibitory effect on cell growth canhowever compensate for this, so that no increase ofmonoclonal antibody titer might be obtained. We showin this article an experiment with addition of butyricacid in the middle of the growth phase of a batchculture, as a strategy to take real profit of such anaddition by a significant increase of final monoclonalantibody concentration. Indeed, in this way asignificant cell density could be obtained before theaddition of butyric acid, while the remaining culturetime was still sufficiently long for its action,resulting in a two fold increase of final monoclonalantibody titer. The experiment was carried out in a 2 L bioreactor, showing the real practical interest ofsuch an addition for the large scale production ofproteins. Furthermore, analysis of the produced IgG bySDS-PAGE and Western blot did not reveal structuralchanges after stimulation by butyric acid. An originalpoint of our study is the characterization of the cellbehaviour, by flow cytometry and other relatedtechniques, leading to a better insight in the effectof the butyric acid addition on cell growth andmonoclonal antibody production. Although there existsa lot of knowledge about the effects of butyrate oncells in the field of molecular biology, our article isat our knowledge one of the first to show some of itseffects on cell behaviour in bioreactor culture,carried out under perfectly defined and controlledconditions, and with the aim to stimulate monoclonalantibody production.
在我们实验室进行的早期实验表明，丁酸对杂交瘤细胞单克隆抗体产生具有刺激作用。然而，它同时对细胞生长的抑制作用可以抵消这一点，因此可能无法获得单克隆抗体滴度的增加。在本文中，我们展示了在分批培养的生长中期添加丁酸的实验，这是一种通过显著增加最终单克隆抗体浓度来真正利用这种添加的策略。事实上，通过这种方式，在添加丁酸之前可以获得显著的细胞密度，而剩余的培养时间仍然足够长，可以发挥其作用，从而使最终单克隆抗体滴度增加一倍。该实验在 2 L 生物反应器中进行，显示了这种添加在大规模生产蛋白质方面的实际应用。此外，通过 SDS-PAGE 和 Western blot 对产生的 IgG 进行分析后，没有发现结构变化。我们研究的一个新颖之处在于通过流式细胞术和其他相关技术对细胞行为进行了表征，从而深入了解丁酸添加对细胞生长和单克隆抗体产生的影响。尽管在分子生物学领域中已经有很多关于丁酸盐对细胞影响的知识，但据我们所知，我们的文章是首次展示其在生物反应器培养中对细胞行为的一些影响的文章之一，该实验在完全定义和控制的条件下进行，并旨在刺激单克隆抗体的产生。
