Equipment for studying protein aggregation

重组蛋白是一类重要的生物药物。由于这些蛋白质不是自然产生的，它们表现出明显的自结合成聚集体的倾向。已知的几个因素，如高浓度、极端的pH、高剪切率和热应力，都会促进聚集体的形成。蛋白质聚集体可以在细胞培养、纯化和储存过程中形成。临床使用的蛋白质应该是无聚集的，因为这些蛋白质是免疫原性的，并已知会在患者中引起不良反应。蛋白质聚集也在ALS、阿尔茨海默病、帕金森病和变异型克雅氏病(VCJD)等人类疾病中发挥关键作用，所有这些疾病都涉及聚集的错误折叠蛋白(AMP)。因此，检测和量化蛋白质聚集体的能力在工业和临床应用中都是重要的。目前使用的分析方法速度慢，效率低。申请者研究小组最近发表的论文描述了基于疏水相互作用膜色谱(HIMC)的快速有效分离和分析蛋白质聚集体的方法。这些方法能够拆分单体、二聚体、三聚体、四聚体和五聚体形式的重组单抗。然而，这些不同形式的身份的确认依赖于缓慢的离线技术，如凝胶电泳法。多角度光散射(或MALS)测量允许快速和在线确定分子大小。拟议购买的MALS探测器将大大提高申请人实验室开发的HIMC技术的能力，并将总体上导致开发出分析蛋白质聚集体的极其有效的方法。

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.