在线高效液相色谱-ICP-质谱法快速标示和定量细胞培养基中的金属成分

IF 3.1 2区 化学 Q2 CHEMISTRY, ANALYTICAL
Cameron J. Stouffer, Sarah K. Wysor and R. Kenneth Marcus
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引用次数: 0

摘要

众所周知,中国仓鼠卵巢(CHO)细胞培养基(CCM)中的金属含量会极大地影响工艺生产率和关键质量属性(CQAs)。金属在 CCM 中以不同的化学形式存在,在单克隆抗体(mAb)生产周期中可能会发生变化。自然,人们推测这些金属的化学形态会影响它们的吸收和新陈代谢。虽然培养基中金属的目标形式是公司指定的,但其商业来源以及有机培养基成分的来源可能会导致浓度和规格与预期配方不同。因此,需要一种方法来确定金属的浓度及其化学形态。本文介绍的是一种利用聚丙烯毛细管通道(C-CP)纤维柱的高效液相色谱法(HPLC)和电感耦合等离子体质谱法(ICP-MS)测定五种目标金属(锰、铁、钴、铜和锌)的方法。采用 50 μL 注入 CCM 上清液的有效定量方法,可确定金属种类以及与报告配方水平的浓度偏差。文中介绍了该方法实用性的两个案例研究:潜在的化学污染和保质期导致的物种变异。该方法的进一步发展与质量控制、污染物鉴定、培养基稳定性/降解产物评估以及生长培养基中全细胞金属吸收测量有关。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

In-line HPLC-ICP-MS method for the rapid speciation and quantification of metal constituents in cell culture media

In-line HPLC-ICP-MS method for the rapid speciation and quantification of metal constituents in cell culture media

In-line HPLC-ICP-MS method for the rapid speciation and quantification of metal constituents in cell culture media

It is well established that the metal content in Chinese hamster ovary (CHO) cell culture media (CCM) greatly affects process productivity and critical quality attributes (CQAs). Metals exist in diverse chemical forms in CCM, which may change during the monoclonal antibody (mAb) production cycle. Naturally, it is postulated that the chemical speciation of these metals affects their uptake and metabolism. While the targeted forms of metals in media are company-specified, their commercial sources, as well as those of the organic media constituents, may result in concentrations and speciation that differ from the intended formulae. Therefore, there is a need for a method to determine the concentration of metals and their chemical forms. Presented here is a methodology to speciate inorganic-versus-ligated metals using high-performance liquid chromatography (HPLC) employing a polypropylene capillary-channeled (C-CP) fiber column with inductively coupled plasma mass spectrometry (ICP-MS) determinations of five target metals (Mn, Fe, Co, Cu, and Zn). A 50 μL injection of CCM supernatant is used for an effective quantification method to identify metal speciation and concentration deviations from reported formula levels. Two case studies as to the utility of the methodology are presented: potential chemical contamination and species variability due to shelf-life. Further development of this method has relevance towards quality control, identification of contaminants, assessment of media stability/degradation products, and measurement of whole cell metal uptake in growth media.

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来源期刊
CiteScore
6.20
自引率
26.50%
发文量
228
审稿时长
1.7 months
期刊介绍: Innovative research on the fundamental theory and application of spectrometric techniques.
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