The impact of microcarrier culture optimization on the glycosylation profile of a monoclonal antibody.
The impact of microcarrier culture optimization on the glycosylation profile of a monoclonal antibody.
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DOI：
10.1186/2193-1801-2-25
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发表时间：
2013-12
期刊：
影响因子：
--
通讯作者：
Azeredo J
中科院分区：
文献类型：
--
作者：
Costa AR;Withers J;Rodrigues ME;McLoughlin N;Henriques M;Oliveira R;Rudd PM;Azeredo J
微载体广泛用于附着依赖性细胞的大规模培养，增加了细胞密度，最终提高了产品产量。在这些过程中，特定的培养条件会影响产品的质量，这与其糖基化模式密切相关。此外，该领域缺乏研究，因此需要更好地了解微载体培养对产品糖基化的影响。因此，在这项工作中，在不同条件下评估了在Cytodex 3中生长的CHO-K1细胞产生的单克隆抗体（mAb）的糖基化谱，并与典型贴壁培养的结果进行了比较。研究发现，微载体培养产生的糖基化谱与t型瓶培养具有不同的特征，半乳糖基化普遍增加，聚焦化水平降低，两者都对单抗活性有潜在的积极影响。唾液酰化也有变化，但没有一般的趋势。该研究随后表明，微载体培养中使用的特定培养条件会影响单抗聚糖谱，并且每种功能元素（半乳糖、核心焦点、唾液酸）都受这些条件的独立影响。特别是，当接种时使用一半体积时，聚焦化大大降低（从79%降至55%），并且在摇瓶培养中观察到唾液化显著降低（从23%降至2%）和糖型异质性（从20降至11），这可能与这些培养容器中实现的细胞密度提高有关。
Microcarriers are widely used for the large-scale culture of attachment-dependent cells with increased cell densities and, ultimately, higher product yield. In these processes, the specific culture conditions can affect the quality of the product, which is closely related to its glycosylation pattern. Furthermore, the lack of studies in the area reinforces the need to better understand the effects of microcarrier culture in product glycosylation. Consequently, in this work, the glycosylation profile of a monoclonal antibody (mAb) produced by adherent CHO-K1 cells grown in Cytodex 3 was evaluated under different conditions, and compared to that obtained of typical adherent cultures. It was found that microcarrier cultures result in a glycosylation profile with different characteristics from T-flask cultures, with a general increase in galactosylation and decrease in fucosylation levels, both with a potentially positive impact on mAb activity. Sialylation also varied but without a general tendency. This study then showed that the specific culture conditions used in microcarrier culture influence the mAb glycan profile, and each functional element (galactose, core fucose, sialic acid) is independently affected by these conditions. In particular, great reductions of fucosylation (from 79 to 55%) were obtained when using half volume at inoculation, and notable decreases in sialylation (from 23 to 2%) and glycoform heterogeneity (from 20 to 11 glycoforms) were observed for shake flask culture, potentially associated with the improved cell densities achieved in these culture vessels.
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DOI：
10.3390/ph3010146
发表时间：
2010-01-12
期刊：
Pharmaceuticals (Basel, Switzerland)
影响因子：
--
作者：
Abès R;Teillaud JL
通讯作者：
Teillaud JL
DOI：
10.1074/mcp.m111.010090
发表时间：
2011-10
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Pucić M;Knezević A;Vidic J;Adamczyk B;Novokmet M;Polasek O;Gornik O;Supraha-Goreta S;Wormald MR;Redzić I;Campbell H;Wright A;Hastie ND;Wilson JF;Rudan I;Wuhrer M;Rudd PM;Josić D;Lauc G
通讯作者：
Lauc G
影响因子：
2.2
作者：
Hu, XW;Xiao, CZ;Li, ZH
通讯作者：
Li, ZH
影响因子：
2.9
作者：
Butler, M;Burgener, A;Coombs, K
通讯作者：
Coombs, K
影响因子：
2.9
作者：
Hodoniczky, J;Zheng, YZ;James, DC
通讯作者：
James, DC