Combined Dynamic Light Scattering and Raman Spectroscopy Approach for Characterizing the Aggregation of Therapeutic Proteins
| Autor: | Stacy M. Kenyon, Steven Blake, Samiul Amin, Wei Qi, Linda H. Kidder, E. Neil Lewis |
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| Rok vydání: | 2014 |
| Předmět: |
Protein Denaturation
Light Pharmaceutical Science Nanotechnology Protein aggregation Spectrum Analysis Raman Stability (probability) Article protein aggregation Analytical Chemistry lcsh:QD241-441 Avian Proteins Protein Aggregates symbols.namesake lcsh:Organic chemistry Dynamic light scattering Enzyme Stability Spectroscopy Fourier Transform Infrared Drug Discovery Animals Transition Temperature Colloids Physical and Theoretical Chemistry Spectroscopy lysozyme Chemistry Organic Chemistry dynamic light scattering self-assembly Hydrogen-Ion Concentration Protein tertiary structure Characterization (materials science) Design for manufacturability Chemistry (miscellaneous) Raman spectroscopy symbols Molecular Medicine Muramidase Chickens |
| Zdroj: | Molecules Molecules, Vol 19, Iss 12, Pp 20888-20905 (2014) Volume 19 Issue 12 Pages 20888-20905 |
| ISSN: | 1420-3049 |
| DOI: | 10.3390/molecules191220888 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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