Biotechnology letters · 2025
Application of aurintricarboxylic acid to boost CHO cell performance.
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Tao Chao, Meng Jizhou, Wen Dan, Dai Jia, Yu Youyou, Wang Yu, Wei Xiangju, Zhao Qing, Sun Ruiqiang, Zhou Hang
Аннотация
PURPOSE: Currently, a robust process using healthy Chinese hamster ovary (CHO) cells, the dominating host for biopharmaceutical protein therapeutics, is crucial for high productivity and successful scaling-up. However, issues which are originated from cell performance and detrimental to protein products such as dramatic viability drop and accumulation of toxic metabolites still exist. Aurintricarboxylic acid (ATA), reported as an antioxidant chelator to prevent apoptosis and boost cell growth, was applied as an additive in this study to address the above problems.
METHODS: Two CHO cells were cultivated by fed-batch culture with ATA of different concentrations and adding strategies supplemented. The reactive oxygen species (ROS) test and confocal microscopy have been used to illustrate the working mechanism of ATA.
RESULTS: ATA was proved to be effective in maintaining viability and improving lactate performance in the late fed-batch culture stage for both tested clones. Briefly, the harvest viability was increased by at least 10% after ATA was introduced, and the suppression of lactate accumulation was observed with ATA addition. Moreover, besides adding ATA in the fed-batch stage, a novel and favorable process to involve ATA in seed train was developed, which improved the performance of seed and further benefitted the cell performance and productivity during fed-batch culture. And the ATA was detected to penetrate into CHO cells and suppress the ROS generation intracellularly.
CONCLUSION: Overall, this study advances current knowledge with respect to ATA's capability of enhancing cell performances for CHO cell manufacturing and introduces a novel process of hyper seed train.
METHODS: Two CHO cells were cultivated by fed-batch culture with ATA of different concentrations and adding strategies supplemented. The reactive oxygen species (ROS) test and confocal microscopy have been used to illustrate the working mechanism of ATA.
RESULTS: ATA was proved to be effective in maintaining viability and improving lactate performance in the late fed-batch culture stage for both tested clones. Briefly, the harvest viability was increased by at least 10% after ATA was introduced, and the suppression of lactate accumulation was observed with ATA addition. Moreover, besides adding ATA in the fed-batch stage, a novel and favorable process to involve ATA in seed train was developed, which improved the performance of seed and further benefitted the cell performance and productivity during fed-batch culture. And the ATA was detected to penetrate into CHO cells and suppress the ROS generation intracellularly.
CONCLUSION: Overall, this study advances current knowledge with respect to ATA's capability of enhancing cell performances for CHO cell manufacturing and introduces a novel process of hyper seed train.