采用中空纤维生物反应器进行高细胞密度杂交瘤培养，对代谢参数进行多元分析和优化抗体生产。

Multivariate analysis of metabolic parameters and optimization of antibody production using high cell density hybridoma in hollow fiber bioreactors.

机构信息

Department of Chemical and Materials Engineering, Chang Gung University, 259, Wen-Hwa First Road, Kwei-Shan, Tao-Yuan 333, Taiwan, ROC.

Research Center for Chinese Herbal Medicine and Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, 261, Wen-Hwa First Road, Taoyuan, Taiwan, ROC.

出版信息

Biotechnol Lett. 2019 Sep;41(8-9):963-977. doi: 10.1007/s10529-019-02712-3. Epub 2019 Jul 19.

DOI:10.1007/s10529-019-02712-3

PMID:31325004

Abstract

OBJECTIVES

The relationships of manipulation of culture temperature and medium circulation rate on the metabolic parameters were regressed by multiple linear regression analysis in hollow fiber bioreactors (HFB).

RESULTS

The high circulation rate could significantly enhance the oxygen consumption of the hybridoma cells and the medium's oxidation-reduction potential. A mildly hypothermic condition of 36 °C and a circulation rate of 182.5 mL/min could support the hybridoma had the maximal antibody titer of 60.75 μg/mL for 20 days. When the ammonium ion was 65 ppm or lactate close to 2.6 g/L, the medium was replaced to maintain the stable and healthy cells at the high cell concentration of 3.33 × 10/mL for continuous antibody production. Two serum-free media could be successfully applied to this perfusion system and maintain hybridoma growth and antibody production.

CONCLUSION

The single-use HFBs could provide the advantages including high cell density, low shear stress, and continuous antibody production.

摘要

目的

通过多元线性回归分析，研究中空纤维生物反应器（HFB）中培养温度和培养基循环率对代谢参数的影响。

结果

高循环率可显著提高杂交瘤细胞的耗氧量和培养基氧化还原电位。36°C 的轻度低温和 182.5 mL/min 的循环率可支持杂交瘤在 20 天内产生最大抗体效价 60.75μg/mL。当氨离子浓度为 65ppm 或乳酸接近 2.6g/L 时，更换培养基以在 3.33×10/mL 的高细胞浓度下维持稳定和健康的细胞，从而实现连续的抗体生产。两种无血清培养基均可成功应用于该灌注系统，维持杂交瘤的生长和抗体的生产。