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不同松针提取物对中国仓鼠卵巢细胞生长和单克隆抗体质量的影响。

Effects of various pine needle extracts on Chinese hamster ovary cell growth and monoclonal antibody quality.

机构信息

Anhui University of Chinese Medicine, Hefei, China.

Yangtze Delta Drug Advanced Research Institute, Nantong, China.

出版信息

Prep Biochem Biotechnol. 2023 Oct;53(9):1081-1091. doi: 10.1080/10826068.2023.2166959. Epub 2023 Feb 9.

Abstract

Chinese hamster ovary (CHO) cells are commonly used as "bio-machines" to pro-duce monoclonal antibodies (mAb) because of their ability to produce very complex proteins. In this study, we evaluated the effects of pine needle water extract (PNWE), pine needle ethanol extract (PNEE), and pine needle polysaccharide extract (PNPE) on the CHO cell growth, mAb production and quality using a Fed-batch culture process. PNPE maintained high VCD and viability, and the titer increase was correlated with its concentration. Three extracts effectively reduced the acidic charge variant and modulated mAb glycosylation. PNPE had the most profound effect, with G0F decreasing by 8.7% and G1Fa increasing by 6.7%. The change in the glycoform was also closely related to the PNPE concentration. This study demonstrated that PNPE could facilitate CHO cell growth, increase the mAb production, decrease acidic charge variants, and regulate mAb glycoforms. To identify the components responsible for the above changes, the sugar and flavonoid contents in the extracts were determined, and the chemical compounds were identified by LC-MS, resulting in 38 compounds identified from PNPE. Rich in sugars and flavonoids in these three extracts may be related to increased CHO cell growth and productivity, and changes in glycoforms.

摘要

中国仓鼠卵巢(CHO)细胞因其能够产生非常复杂的蛋白质而被广泛用作生产单克隆抗体(mAb)的“生物机器”。在这项研究中,我们使用分批补料培养工艺评估了松针水提物(PNWE)、松针乙醇提物(PNEE)和松针多糖提物(PNPE)对 CHO 细胞生长、mAb 生产和质量的影响。PNPE 保持了较高的 VCD 和活力,效价的增加与浓度相关。三种提取物有效地减少了酸性电荷变异体并调节了 mAb 的糖基化。PNPE 的影响最为显著,G0F 降低了 8.7%,G1Fa 增加了 6.7%。糖型的变化也与 PNPE 浓度密切相关。本研究表明,PNPE 可以促进 CHO 细胞生长,增加 mAb 产量,减少酸性电荷变异体,并调节 mAb 糖型。为了确定导致上述变化的成分,测定了提取物中的糖和类黄酮含量,并通过 LC-MS 鉴定了化合物,从 PNPE 中鉴定出 38 种化合物。这些提取物中富含糖和类黄酮,可能与 CHO 细胞生长和产率的增加以及糖型的变化有关。

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