Zeeshan, Farrukh and Tabbassum, Misbah and Jorgensen, Lene and Medlicott, Natalie J. (2018) Investigation on Secondary Structure Perturbations of Proteins Embedded in Solid Lipid Matrices as a Novel Indicator of their Biological Activity upon In Vitro Release. AAPS PharmSciTech, 19 (2). pp. 769-782. ISSN 1530-9932, DOI https://doi.org/10.1208/s12249-017-0883-1.
Full text not available from this repository.Abstract
Protein biologics are prone to conformational changes during formulation development. Limited methods are available for conformational analysis of proteins in solid state and in the presences of formulation excipients. The aim of this study was to investigate the secondary structures of proteins encased in solid lipid matrices as a novel indicator of their stability upon in vitro release. Model proteins namely catalase and lysozyme were incorporated into lipid namely Precirol® AT05 (glycerol palmitostearate, melting point 58°C) at 30% w/w loading using melting and mixing and wet granulation methods. Attenuated total reflectance (ATR-FTIR) spectroscopy, size-exclusion chromatography (SEC) and biological activity analyses were performed. The information about secondary structure was acquired using second derivative analysis of amide-I band (1600–1700 cm−1). ATR analysis demonstrated interference of lipid spectrum with protein amide-I band which was subsequently subtracted to allow the analysis of the secondary structure. ATR spectra amide-I bands showed shifts peak band positions compared to native protein for matrices prepared using wet granulation. SEC analysis gave evidence of protein aggregation for catalase which was increased using wet granulation. The biological activity of catalase was statistically different from that of control and was affected by the incorporation method and was found to be in alignment with ATR spectral changes and extent of aggregation. In conclusion, ATR spectroscopy could analyze protein secondary structure in lipid matrices provided lipid interference was minimized. The ATR spectral changes and formation of aggregates can indicate the loss in biological activity of protein released from solid lipid matrices.
| Item Type: | Article |
|---|---|
| Funders: | University of Otago, Dunedin, New Zealand |
| Additional Information: | Misbah Tabbassum. Master Degree. Department of Chemistry, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, MALAYSIA |
| Uncontrolled Keywords: | ATR spectroscopy; Biological activity; Lipid matrices; Proteins; Size-exclusion chromatography |
| Subjects: | Q Science > QD Chemistry R Medicine > RS Pharmacy and materia medica |
| Divisions: | Faculty of Science > Department of Chemistry |
| Depositing User: | Ms. Juhaida Abd Rahim |
| Date Deposited: | 28 Jan 2019 02:03 |
| Last Modified: | 28 Jan 2019 02:03 |
| URI: | http://eprints.um.edu.my/id/eprint/20175 |
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