Combined Dynamic Light Scattering and Raman Spectroscopy Approach for Characterizing the Aggregation of Therapeutic Proteins

Molecules

E. Neil Lewis¹  Wei Qi²  Linda H. Kidder²  Samiul Amin²  Stacy M. Kenyon²  Steven Blake² 

[1] Malvern Biosciences Inc., 7221 Lee Deforest Drive, Suite 300, Columbia, MD 21046, USA;

关键词: dynamic light scattering;    Raman spectroscopy;    lysozyme;    protein aggregation;    self-assembly;   

DOI  :  10.3390/molecules191220888

来源: mdpi

【 摘 要 】

Determination of the physicochemical properties of protein therapeutics and their aggregates is critical for developing formulations that enhance product efficacy, stability, safety and manufacturability. Analytical challenges are compounded for materials: (1) that are formulated at high concentration, (2) that are formulated with a variety of excipients, and (3) that are available only in small volumes. In this article, a new instrument is described that measures protein secondary and tertiary structure, as well as molecular size, over a range of concentrations and formulation conditions of low volume samples. Specifically, characterization of colloidal and conformational stability is obtained through a combination of two well-established analytical techniques: dynamic light scattering (DLS) and Raman spectroscopy, respectively. As the data for these two analytical modalities are collected on the same sample at the same time, the technique enables direct correlation between them, in addition to the more straightforward benefit of minimizing sample usage by providing multiple analytical measurements on the same aliquot non-destructively. The ability to differentiate between unfolding and aggregation that the combination of these techniques provides enables insights into underlying protein aggregation mechanisms. The article will report on mechanistic insights for aggregation that have been obtained from the application of this technique to the characterization of lysozyme, which was evaluated as a function of concentration and pH.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.