进料速度对鼠杂交瘤细胞补料分批培养中生长速率和抗体产量的影响。

Effect of feed rate on growth rate and antibody production in the fed-batch culture of murine hybridoma cells.

机构信息

Department of Chemical Engineering, Sydney University, Sydney, NSW, 2006, Australia.

出版信息

Cytotechnology. 2000 Mar;32(3):229-42. doi: 10.1023/A:1008169417980.

DOI:10.1023/A:1008169417980

PMID:19002984

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3449896/

Abstract

Batch and fed-batch cultures of a murine hybridomacell line (AFP-27) were performed in a stirred tankreactor to estimate the effect of feed rate on growthrate, macromolecular metabolism and antibodyproduction. Macromolecular composition was foundto change dynamically during batch culture ofhybridoma cells possibly due to active production ofDNA, RNA and protein during the exponential phase.Antibody synthesis is expected to compete with theproduction of cellular proteins from the amino acidpool. Therefore, it is necessary to examine therelationship between cell growth in terms of cellularmacromolecules and antibody production. In this study,we searched for an optimum feeding strategy bychanging the target specific growth rate in fed-batchculture to give higher antibody productivity whileexamining the macromolecular composition. Concentratedglucose (60 mM) and glutamine (20 mM) in DR medium(1:1 mixture of DMEM and RPMI) with additional aminoacids were fed continuously to the culture and thefeed rate was updated after every sampling to ensureexponential feeding (or approximately constantspecific growth rate). Specific antibody productionrate was found to be significantly increased in thefed-batch cultures at the near-zero specific growthrate in which the productions of cellular DNA, RNA,protein and polysaccharide were strictly limited byslow feeding of glucose, glutamine and other nutrients. Possible implications of these results are discussed.

摘要

在搅拌罐反应器中进行了鼠杂交瘤细胞系（AFP-27）的分批和补料分批培养，以估计补料率对生长速率、生物大分子代谢和抗体产生的影响。在杂交瘤细胞的分批培养过程中，生物大分子组成被发现是动态变化的，这可能是由于在指数生长期细胞主动合成 DNA、RNA 和蛋白质所致。抗体的合成预计会与从氨基酸池中产生细胞蛋白质竞争。因此，有必要根据细胞的生物大分子和抗体产生来检查细胞生长之间的关系。在这项研究中，我们通过改变补料分批培养中的目标比生长速率，搜索了一种最佳的补料策略，以在考察生物大分子组成的同时，获得更高的抗体生产力。浓缩葡萄糖（60mM）和谷氨酰胺（20mM）在 DR 培养基（DMEM 和 RPMI 的 1:1 混合物）中，并添加额外的氨基酸，连续补加到培养物中，并且在每次采样后更新补料率，以确保指数补料（或近似恒定的比生长速率）。在接近零比生长速率的补料分批培养中，细胞 DNA、RNA、蛋白质和多糖的产生受到葡萄糖、谷氨酰胺和其他营养物质的缓慢补料的严格限制，发现特异性抗体的产生速率显著提高。讨论了这些结果的可能意义。