Manufacturing Sciences & Technology, Bristol-Myers Squibb Company, 6000 Thompson Road, East Syracuse, NY 13057, USA.
Biotechnol Prog. 2011 Mar-Apr;27(2):442-50. doi: 10.1002/btpr.568. Epub 2011 Mar 1.
The use of yeast- and plant-derived hydrolysates in cell culture production processes has sparked concerns over the potential immunogenicity risk posed by β-glucans and yeast peptides contained in these raw materials. This article utilizes a combination of in-process testing from large-scale manufacturing and scale-down spiking studies to demonstrate the clearance of β-glucans and yeast peptides through chromatographic steps in the downstream purification process for a monoclonal antibody. β-Glucans were found to flow through most all three modes of chromatography (Protein A, cation and anion exchange) without binding to the resins or the product. Protein A affinity chromatography was found to provide the best clearance factor. The efficacy of the resin sanitization and storage procedures to prevent carryover from one run to the next was also demonstrated. Yeast peptides were found to be metabolized during the cell culture process and were undetectable after the Protein A purification step. The data presented here serve to allay concerns about the use of hydrolysates in cell culture production. The methodology presented here provides a template to demonstrate clearance of β-glucans and yeast peptides through chromatographic steps in downstream processing.
酵母和植物来源的水解产物在细胞培养生产工艺中的应用引发了人们对于这些原材料中β-葡聚糖和酵母肽可能带来的免疫原性风险的关注。本文采用大规模生产过程中的中间测试和缩小规模的添加研究相结合的方法，证明了单克隆抗体下游纯化过程中通过色谱步骤清除β-葡聚糖和酵母肽。β-葡聚糖被发现可以通过大多数三种色谱模式（Protein A、阳离子和阴离子交换）而不与树脂或产品结合。发现 Protein A 亲和色谱提供了最佳的清除因子。还证明了树脂消毒和储存程序可有效防止从一次运行到下一次运行的残留。酵母肽在细胞培养过程中被代谢，在 Protein A 纯化步骤后无法检测到。本文提供的数据有助于减轻人们对细胞培养生产中使用水解产物的担忧。本文提出的方法为通过下游处理中的色谱步骤清除β-葡聚糖和酵母肽提供了模板。
