Effects of Dioscorea opposita Extract on CHO Cell Growth and Monoclonal Antibody Expression

doi:10.19586/j.2095-2341.2023.0020

Current Biotechnology ›› 2023, Vol. 13 ›› Issue (3): 449-456.DOI: 10.19586/j.2095-2341.2023.0020

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Effects of Dioscorea opposita Extract on CHO Cell Growth and Monoclonal Antibody Expression

Li CAO¹^,²(), Shun LUO¹^,³(), Shihai XING¹, Jinshu QIU³, Zhiyong LIN³, Jun LIN³, Xu MENG³, Feng LIU³

^1.School of Pharmacy，Anhui University of Chinese Medicine，Hefei 230012，China
^2.Yangtze Delta Drug Advanced Research Institute，Jiangsu Nantong 226133，China
^3.Thousand Oaks Biopharmaceuticals Co. Ltd. ，Jiangsu Nantong 226133，China

Received:2023-02-22 Accepted:2023-03-14 Online:2023-05-25 Published:2023-06-12
Contact: Shun LUO

山药提取物对CHO细胞生长及抗体表达的影响

曹丽¹^,²(), 罗顺¹^,³(), 邢世海¹, 仇金树³, 蔺智勇³, 林军³, 孟旭³, 刘峰³

^1.安徽中医药大学药学院，合肥 230012
^2.南通市海门长三角药物高等研究院，江苏南通 226133
^3.澳斯康生物（南通）股份有限公司，江苏南通 226133

通讯作者: 罗顺
作者简介:曹丽 E-mail: 2279763127@qq.com；
基金资助:
安徽省高校科学研究项目(YJS20210493);安徽省高校自然科学研究重点项目(KJ2020A0387)

Abstract

Abstract:

The aim of this study was to investigate the effect of Chinese yam extract （DOE） on CHO cell growth and antibody expression. By adding different concentrations of DOE， the cell density， cell viability， the expression of antibody in cell supernatant， and the content of single antibody were measured respectively. Meanwhile， the antibody charge variants were detected and the glycosyl composition and relative content of antibody and the relative binding activity of protein were analyzed. In addition， the activities of malondialdehyde （MDA） and superoxide dismutase （SOD） in cells were measured during cell growth. The results showed that 5 mg·mL^-1 of DOE could significantly promote the growth of CHO cells， increase the density of living cells， cell viability and SOD activity， and reduce the content of MDA， and also increase the content of intracellular antibodies and monoclonal antibodies. The addition of DOE could change the content of some sugar types of antibody to a certain extent. With the increase of DOE concentration， the relative content of G0F and Man5 showed a trend of first decreasing and then slowly increasing， while the content of G1F sugar type gradually increased， and the change of G0 was relatively stable and small. After the addition of DOE， there was no significant difference in the content of acid charge variant， main peak and basic charge variant in the antibody （P>0.05）， and the relative binding activity of the antibody remained between 102% and 135% of the reference sample. DOE did not affect the binding activity of the antibody. Overall， DOE played an important role in cell growth and antibody expression during cell culture. The addition of 5 mg·mL^-1 of DOE had the best effect on cell growth and antibody expression. The result provided a reference for the future study of the effects of Chinese herbal extracts on CHO cell growth and antibody expression.

Key words: Chinese yam extract, CHO cells, cell growth, antibodies, oxidative stress

摘要：

探究山药提取物（Dioscorea oppsita extract，DOE）对中国仓鼠卵巢细胞（Chinese hamster ovary cells，CHO）生长及抗体表达的影响。通过添加不同浓度的DOE，采用流加培养的方式，分别测定细胞密度、细胞活力、细胞上清中抗体表达量、单抗体含量，检测抗体电荷变体并分析抗体的糖基组成及相对含量和蛋白相对结合活性，以及在细胞生长过程中测定细胞内丙二醛（malondialdehyde，MDA）和超氧化物歧化酶（superoxide dismutase，SOD）的活性。结果表明，5 mg·mL^-1 DOE可以明显地促进CHO细胞的生长，提高活细胞密度、细胞活力和SOD活性并降低MDA含量、提高细胞内抗体和单抗含量。添加DOE在一定程度上能够改变抗体某些糖型含量，且随着DOE浓度的增加，糖型G0F和Man5相对含量呈现先降低后缓慢增加的趋势，G1F糖型含量则逐渐升高，G0则变化较小相对较稳定。添加DOE后，抗体中酸性电荷变体、主峰和碱性电荷变体含量均不存在统计学差异（P>0.05），其抗体相对结合活性均保持在参比品的102%~135%之间，DOE不影响抗体的结合活性。综上所述DOE在细胞培养过程中对细胞的生长、抗体的表达方面发挥着重要作用，添加5 mg·mL^-1 DOE对细胞生长的促进作用和抗体表达效果最佳，研究结果为中药提取物对CHO细胞的生长及抗体表达方面的研究提供了参考。

关键词: 山药提取物, CHO细胞, 细胞生长, 抗体, 氧化应激

CLC Number:

R285.5

Li CAO, Shun LUO, Shihai XING, Jinshu QIU, Zhiyong LIN, Jun LIN, Xu MENG, Feng LIU. Effects of Dioscorea opposita Extract on CHO Cell Growth and Monoclonal Antibody Expression[J]. Current Biotechnology, 2023, 13(3): 449-456.

曹丽, 罗顺, 邢世海, 仇金树, 蔺智勇, 林军, 孟旭, 刘峰. 山药提取物对CHO细胞生长及抗体表达的影响[J]. 生物技术进展, 2023, 13(3): 449-456.

Figures/Tables 8

Fig. 1 Viable cell densityand and cell viability of CHO cells under different DOE concentrations

Fig. 2 Changes of intracellular MDA content

Fig. 3 Changes of intracellular SOD activity

Fig. 4 The mAb yield of CHO cells under different concentrations of DOE

Fig. 5 Content of mAb monomers in CHO cells at different DOE concentrations

Fig. 6 N-glycosylation modification of CHO cells under the action of DOE at different concentrations

Fig. 7 The mAb charge variant content produced by CHO cells at different DOE concentrations

Table 1 Relative binding activity under different DOE concentrations

组别	相对结合活性/%	RSD/%
CON	121.39±7.56	2.75
SY-1	135.03±6.52	3.21
SY-5	125.86±10.31	1.27
SY-8	117.84±8.27	4.35
SY-10	116.48±9.54	3.78
SY-15	102.88±8.76	2.86

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Effects of Dioscorea opposita Extract on CHO Cell Growth and Monoclonal Antibody Expression

山药提取物对CHO细胞生长及抗体表达的影响

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