A new microfluidic tool for rapid analysis of protein stability and integrity in bioprocesses

一种新的微流体工具，用于快速分析生物过程中蛋白质的稳定性和完整性

• 批准号: BB/E005942/1
• 负责人: Paul Dalby
• 金额: $ 54.05万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FE005942%2F1
• 关键词: new; microfluidic; tool; rapid; analysis

Analysis of protein stability is currently too slow and requires too much of an exceedingly valuable biopharmaceutical to be useful in guiding bioprocess development or control. Introducing the first microfluidic method for protein stability testing will reduce sample use and cost of analysis by up to 108-fold over microwell-based analysis. Combined expertise from biochemical engineering and the London Centre for Nanotechnology will enable this analysis with parallelism for up to 1000 samples per day. The new generation of protein-based medicines has rapidly become a $30billion-a-year industry addressing previously untreatable diseases. They have the potential for much further growth but a principal constraint is the high cost of the manufacturing methods required to preserve the structural integrity of proteins with limited stability. The ability to perform rapid and parallel protein stability characterisation experiments, at the microfluidic scale, is essential to enable: a) the rapid optimisation of therapeutic protein formulations; and b) the real-time monitoring of protein product quality in process-, microwell- and microfluidic scale bioprocess development experiments. Our preliminary research has demonstrated protein stability determination using fluorescence measurements at the microwell scale (Aucamp et al., 2005). The aims of this proposal are to a) explore the fundamentals that impact on measurement accuracy and sensitivity at the microfluidic scale, so as to significantly decrease the sample volumes required for protein stability measurement; b) establish a microfluidic denaturation technique; c) overcome the challenges that will enable broad application to bioprocessing and formulation of biopharmaceutical protein products.

目前，蛋白质稳定性分析速度太慢，并且需要太多极其有价值的生物制药才能用于指导生物过程开发或控制。引入第一个用于蛋白质稳定性测试的微流体方法将比基于微孔的分析减少多达 108 倍的样品使用和分析成本。生物化学工程和伦敦纳米技术中心的专业知识相结合，可以实现每天最多 1000 个样本的并行分析。新一代蛋白质药物已迅速成为一个年产值 300 亿美元的产业，用于解决以前无法治疗的疾病。它们具有进一步增长的潜力，但主要限制是保持蛋白质结构完整性和稳定性有限所需的制造方法成本高昂。在微流体规模上进行快速、并行的蛋白质稳定性表征实验的能力对于实现以下目标至关重要：a) 治疗性蛋白质配方的快速优化； b) 在工艺、微孔和微流体规模生物工艺开发实验中实时监测蛋白质产品质量。我们的初步研究已经证明使用微孔规模的荧光测量来确定蛋白质稳定性（Aucamp 等人，2005）。该提案的目的是a）探索影响微流体尺度测量精度和灵敏度的基本原理，从而显着减少蛋白质稳定性测量所需的样品体积； b) 建立微流体变性技术； c) 克服挑战，使生物制药蛋白产品的生物加工和配方得到广泛应用。

项目成果

Optically induced thermal gradients for protein characterization in nanolitre-scale samples in microfluidic devices.

• DOI: 10.1038/srep02130
• 发表时间: 2013
• 期刊: SCIENTIFIC REPORTS
• 影响因子: 4.6
• 作者: Sagar, D. M.;Aoudjane, Samir;Gaudet, Matthieu;Aeppli, Gabriel;Dalby, Paul A.
• 通讯作者: Dalby, Paul A.

Virus lasers for biological detection

用于生物检测的病毒激光器

• DOI: 10.3929/ethz-b-000359804
• 发表时间: 2019
• 作者: Hales, John E.

Proof-of-concept analytical instrument for label-free optical deconvolution of protein species in a mixture.

用于混合物中蛋白质种类的无标记光学解卷积的概念验证分析仪器。

• DOI: 10.1016/j.chroma.2021.461968
• 发表时间: 2021
• 期刊: Journal of chromatography. A
• 作者: Hales JE

High-Throughput Characterization of Proteins From Microtiter Plates to Microfluidics

从微量滴定板到微流控的蛋白质高通量表征

• 发表时间: 2009
• 作者: Dalby PA