Protein-poly(amino acid) precipitation stabilizes a therapeutic protein l-asparaginase against physicochemical stress
| Autor: | Takuya Maruyama, Takaaki Kurinomaru, Kenji Handa, Izaki Shunsuke, Kentaro Shiraki, Tomoaki Kimoto |
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| Rok vydání: | 2015 |
| Předmět: |
Hot Temperature
genetic structures Drug Storage Bioengineering behavioral disciplines and activities Applied Microbiology and Biotechnology L asparaginase Stress (mechanics) Electrolytes Motion Asparaginase Chemical Precipitation Polylysine chemistry.chemical_classification Chromatography Aqueous solution biology Precipitation (chemistry) Chemistry Protein Stability Therapeutic protein Enzyme assay Amino acid Enzyme Activation Solutions Solubility biology.protein Protein concentration Oxidation-Reduction psychological phenomena and processes Biotechnology |
| Zdroj: | Journal of bioscience and bioengineering. 120(6) |
| ISSN: | 1347-4421 |
| Popis: | Long-term storage in aqueous solution has been demanded for the practical application of therapeutic proteins. Recently, a precipitation-redissolution method was proposed to prepare salt-dissociable protein-polyelectrolyte complex (PPC). To elucidate the utility of the complex for storage of proteins, we investigated the stress tolerance of PPC precipitates containing l-asparaginase (ASNase) and poly-l-lysine (polyK). PPC precipitate containing ASNase and polyK was prepared by precipitation-redissolution method. The sample was treated to three types of stress, i.e., heat, shaking, and oxidation. The protein concentration, enzyme activity, and CD spectrum of the supernatants of samples were measured after stressed. PPC precipitate consisting of ASNase and polyK showed tolerance against thermal and shaking stress compared to the native solution. In addition, PPC precipitate protected ASNase from inactivation by oxidation. PPC precipitate of ASNase/polyK complex successfully stabilized ASNase against physicochemical stresses. These results suggest that the PPC precipitate has great potential as a storage method in aqueous solution for unstable proteins. |
| Databáze: | OpenAIRE |
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