Predictive approach for protein aggregation: Correlation of protein surface characteristics and conformational flexibility to protein aggregation propensity
| Autor: | Lara Galm, Sven Amrhein, Jürgen Hubbuch |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Ammonium sulfate
Aqueous solution Hofmeister series Bioengineering 02 engineering and technology Protein aggregation 021001 nanoscience & nanotechnology 030226 pharmacology & pharmacy Applied Microbiology and Biotechnology 03 medical and health sciences chemistry.chemical_compound Molecular dynamics Crystallography 0302 clinical medicine Protein structure chemistry Biophysics Lysozyme 0210 nano-technology Protein secondary structure Biotechnology |
| Zdroj: | Biotechnology and Bioengineering. 114:1170-1183 |
| ISSN: | 0006-3592 |
| DOI: | 10.1002/bit.25949 |
| Popis: | The aggregation of proteins became one of the major challenges in the development of biopharmaceu-ticals since the formation of aggregates can affect drug quality and immunogenicity. However, aggregation mechanisms are highly complex and the investigation requires cost, time, and material intensive experi-mental effort. In the present work, the predictive power of protein characteristics for the phase behavior of three different proteins which are very similar in size and structure was studied. In particular, the surface hydrophobicity, zeta potential, and conformational flexibility of human lysozyme, lysozyme from chicken egg white, and α-lactalbumin at pH 3, 5, 7, and 9 were assessed and examined for correlation with experimental stability studies focusing on protein phase behavior induced by sodium chloride and ammonium sulfate. The molecular dynamics (MD) simulation based study of the conformational flexibility without precipitants was able to identify highly flexible protein regions which could be associated to the less regular secondary structure elements and random coiled and terminal regions in particular. Conformational flex-ibility of the entire protein structure and protein surface hydrophobicity could be correlated to differing aggregation propensities among the studied proteins and could be identified to be applicable for predic-tion of protein phase behavior in aqueous solution without precipitants. For prediction of protein phase behavior and aggregation propensity in aqueous solution with precipitants, protein flexibility was further studied in dependency of salt concentration and species by means of human lysozyme. Even though the results of the salt dependent MD simulations could not be shown to be sufficient for prediction of salt depending phase behavior, this study revealed a more pronounced destabilizing effect of ammonium sulfate in comparison to sodium chloride and thus, was found to be in good agreement with theoretical considerations along the Hofmeister series as well as experimentally evaluated phase behavior. Biotechnol. Bioeng. 2017;114: 1170-1183. © 2016 Wiley Periodicals, Inc. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text