| Preparation and characterisation of spray-dried and crystallised trypsin: FT-Raman study to detect protein denaturation after thermal stress. | |
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MedLine Citation:
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PMID: 17236751 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The production of stable protein formulations is difficult due to unique properties of proteins. Accordingly, spray drying and crystallisation techniques were assessed for their effects on trypsin, a model protein. Samples were investigated using polarising microscopy, thermogravimetry, differential scanning calorimetry (DSC), FT-Raman spectroscopy and enzymatic assay. Unprocessed, spray-dried and crystallised trypsin were evaluated in solution for secondary structure in low and high protein concentrations using aqueous state FT-Raman spectroscopy and for folding reversibility employing high sensitivity differential scanning calorimetry. Spray-dried trypsin showed FT-Raman spectral changes and less biological activity, after rehydration, compared with unprocessed and crystallised trypsin. Crystals maintained activity better than did the spray-dried form and retained a higher folding reversibility compared to unprocessed and spray-dried protein. Proteins may denature with structural changes under thermal stress and lose their activities. Thus, this research studied the effect of heating solid unprocessed, spray-dried and crystallised trypsin samples on their secondary structures, using FT-Raman spectroscopy, to identify the influence of the initial solid form on its propensity for thermal denaturation and whether this can be correlated with catalytic activity. DSC heated protein samples to two temperatures, one before the apparent denaturation temperature (T(m)) and the other after the T(m). Samples heated below their T(m) showed some perturbations of the secondary structure and some activity, whilst materials rose to the higher temperature were insoluble with complete loss of activity. |
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Authors:
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Robert T Forbes; Brian W Barry; Amal A Elkordy |
Publication Detail:
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Type: Journal Article Date: 2006-12-10 |
Journal Detail:
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Title: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences Volume: 30 ISSN: 0928-0987 ISO Abbreviation: Eur J Pharm Sci Publication Date: 2007 Mar |
Date Detail:
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Created Date: 2007-02-26 Completed Date: 2007-05-23 Revised Date: 2008-11-21 |
Medline Journal Info:
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Nlm Unique ID: 9317982 Medline TA: Eur J Pharm Sci Country: Netherlands |
Other Details:
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Languages: eng Pagination: 315-23 Citation Subset: IM |
Affiliation:
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Drug Delivery Group, School of Pharmacy, University of Bradford, Bradford BD7 1DP, UK. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Calorimetry, Differential Scanning Catalysis Crystallization Desiccation Disulfides / chemistry Fourier Analysis Hot Temperature Microscopy, Polarization Protein Denaturation* Spectrum Analysis, Raman Thermogravimetry Trypsin / chemistry* Vibration |
| Chemical | |
Reg. No./Substance:
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0/Disulfides; EC 3.4.21.4/Trypsin |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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