生产用细胞培养
细胞培养可用于生产各类生物制剂,如疫苗、治疗性蛋白质和抗体以及细胞治疗产品。随着生物疗法和疫苗的需求日益增长,如何通过以下手段提高细胞培养工艺效率越发受到广泛关注:
- 可提高产量的细胞系和细胞培养体系
- 针对扩增和生产优化的细胞培养基
- 有助改善细胞生长和可扩展性的生物反应器
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生产用细胞培养
尽管细菌、酵母和昆虫细胞表达系统也具有重组蛋白过表达能力,但哺乳动物细胞因其具有传播人类病毒、表达单克隆抗体以及引入糖基化等对生产有效生物制品至关重要的翻译后修饰的能力,更为生物制品生产常用。现已开发并使用多种哺乳动物细胞系来生产高水平的治疗性蛋白和抗体,包括CHO、淋巴瘤(NS0,SP2 / 0)和人胚肾(HEK)细胞等细胞系。随着CRISPR-Cas系统等基因编辑技术的不断发展,更可通过改良宿主细胞基因组操纵细胞生长和凋亡,促进基因产物的翻译后修饰,从而进一步提高生产效率和产品质量。
生物制剂生产用培养基
培养基优化是开发生物制品生产细胞培养工艺的另一关键环节。生物制品生产常采采用不含血清和动物来源成分的合成型细胞培养基,目的是减少成分不明确培养基和补剂固有的差异因子,消除工艺污染物和外源性物质,并降低非预期/不良效应风险。由于细胞系各异,需要不同的营养成分才能茁壮生长,因此培养生物体和蛋白的产量很大程度依赖使用的培养基。
生物反应器是上游工艺和生产所采用的大规模培养设备,在扩展扩大生产用细胞培养体系过程中起关键作用。常用的接种培养细胞扩增工艺要求先解冻冻存的细胞,随后经连续扩增后转移到更大规模的培养容器中,如烧瓶、旋转器和搅拌式生物反应器等。达到理想的培养物体积和细胞密度后,将培养体系转移至生产用生物反应器中。生物反应器可提供受控的微环境和营养成分输送途径,从而调节细胞的生长和分化,提高标准化程度和可重现性。尽管采用一次性生物反应器能够提高灵活性、缩短传统固定式设备的清洁和消毒所需的停机时间,但生产过程仍主要采用不锈钢搅拌釜式生物反应器。
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