Equipment for studying protein aggregation

研究蛋白质聚集的设备

• 批准号: 390359-2010
• 负责人: Ghosh, Raja
• 金额: $ 2.56万
• 依托单位: McMaster University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2009
• 资助国家: 加拿大
• 起止时间: 2009-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=443172
• 关键词: Equipment; studying; protein; aggregation

Recombinant proteins comprise an important class of biopharmaceuticals. Since these proteins do not occur naturally, they show significant tendency to self-associate into aggregates. Several factors such as high concentration, extremes of pH, high shear rates and thermal stress are known to encourage aggregate formation. Protein aggregates can be formed during cell culture, purification and storage. Clinically used proteins should be aggregate free since these are immunogenic and are known to cause adverse reactions in patients. Protein aggregation also plays a key role in human diseases such as ALS, Alzheimer's disease, Parkinson's disease and variant Creutzfeldt - Jakob disease (vCJD), all of which involve aggregated misfolded proteins (AMP). The ability to detect and quantify protein aggregates is therefore important in both industrial and clinical applications. Currently used analytical methods are slow and inefficient. Recent papers from the applicants research group have described hydrophobic interaction membrane chromatography (or HIMC) based methods for rapid and efficient separation and analysis of protein aggregates. These method were able to resolve monomer, dimer, trimer, tetramer and pentamer forms of recombinant monoclonal antibodies. However, the confirmation of identity of these different forms relied on slow off-line techniques such as gel electrophoresis. Multi angle light scattering (or MALS) measurement allows rapid and online determination of molecular size. The proposed purchase of a MALS detector would significantly enhance the capability of the HIMC techniques developed in the applicants lab and would overall lead to the development of extremely efficient methods for analyzing protein aggregates.

重组蛋白包括一类重要的生物药物。由于这些蛋白质不是天然存在的，它们显示出显著的自缔合成聚集体的趋势。已知若干因素如高浓度、极端pH、高剪切速率和热应力促进聚集体形成。蛋白质聚集体可以在细胞培养、纯化和储存期间形成。临床使用的蛋白质应该是无聚集体的，因为这些蛋白质具有免疫原性，并且已知会在患者中引起不良反应。蛋白质聚集还在人类疾病如ALS、阿尔茨海默病、帕金森病和变异型克雅氏病（vCJD）中起关键作用，所有这些疾病都涉及聚集的错误折叠蛋白（AMP）。因此，检测和定量蛋白质聚集体的能力在工业和临床应用中都很重要。目前使用的分析方法是缓慢和低效的。来自申请人研究小组的最近的论文描述了用于快速和有效分离和分析蛋白质聚集体的基于疏水相互作用膜色谱（或HIMC）的方法。这些方法能够分辨重组单克隆抗体的单体、二聚体、三聚体、四聚体和五聚体形式。然而，这些不同形式的同一性的确认依赖于缓慢的离线技术，如凝胶电泳。多角度光散射（或MALS）测量允许快速和在线测定分子大小。拟议购买的MALS检测器将显著增强申请人实验室开发的HIMC技术的能力，并将总体上导致开发用于分析蛋白质聚集体的极其有效的方法。