蛋白质表达和生物活性生长因子的净化,用于细胞培养和细胞农业
Cameron Semper1, Alexei Savchenko2
1Department of Microbiology, Immunology and Infectious Disease, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada.
STAR protocols
|June 14, 2023
概括
这项研究引入了一种具有成本效益的生产生物活性生长因子的方法,这对于无血清细胞培养至关重要. 精简的工作流大大降低了生物制药研究中菌生长因子的费用.
科学领域:
- 生物技术是生物技术.
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- 线粒体生长因子是无血清细胞培养基中的关键成分.
- 这些增长因素代表着相当大的成本,高达媒体总支出的95%.
- 高昂的生产成本限制了它们在研究和生物制造中的广泛应用.
研究的目的:
- 为生产生物活性生长因子开发一种通用,低成本的工作流程.
- 能够有效地产生关键生长因子,如基本纤维细胞生长因子和转化生长因子β1.1.
- 为各种生长因子家族提供适合各种生长因子家族的可扩展方法.
主要方法:
- 建立了一个简化的工作流程,包括克隆,表达测试,蛋白质净化和生物活性查.
- 该协议被优化为产生特定的生长因子,包括基本纤维细胞生长因子和转化生长因子β1.1.
- 一般化程序允许微小的修改以适应不同的生长因子家族.
主要成果:
- 开发的工作流程成功地以显著降低的成本产生了生物活性生长因子.
- 纯化的生长因子显示出适用于细胞培养应用的生物活性.
- 该协议在产生基本纤维细胞生长因子和转化生长因子β1.1方面都被证明是有效的.
结论:
- 一种具有成本效益和通用的生产生物活性生长因子的方法已经成功开发出来.
- 这一工作流解决了无血清介质的主要成本驱动因素,使必要的增长因素更容易获得.
- 生产的生长因子具有生物活性,可以直接应用于细胞培养,支持生物技术和生物制药研究的进步.
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