کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6491254 | 43404 | 2015 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Non-chromatographic preparative purification of enhanced green fluorescent protein
ترجمه فارسی عنوان
خالص سازی فرآوری غیر کروماتوگرافی از پروتئین فلورسنت سبز افزایش یافته است
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کلمات کلیدی
تخمیر هموژنیزه شده بوجان، پروتئین فلورسنت سبز پیشرفته کریستالیزاسیون آماده سازی، تصفیه پروتئین، کریستالیزر مخلوط مخلوط،
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
چکیده انگلیسی
Recombinant enhanced green fluorescent protein (eGFP) is used as a marker in numerous applications in biomedical research and diagnostics. For these applications, the macromolecule needs to be provided in a highly purified form. The conventional purification process of eGFP usually consists of multiple subsequent preparative chromatography steps. Since this procedure is costly and time-consuming, an alternative chromatography-free purification process was investigated. This process was a combination of three-phase partitioning (TPP) and preparative crystallization including an ultrafiltration/diafiltration (UF/DF) intermediate step. After the TPP step, eGFP with a purity level suitable for preparative crystallization of 82.5-85.0% and a yield of 84-92% was obtained depending on the scale. After cross-flow UF/DF, the crystallization was performed in parallelized mL-scale stirred tanks. A favorable robust crystal morphology was obtained combined with fast crystallization kinetics when two polyethylenglycols and ethanol were used simultaneously as crystallization additives. The crystallization process can easily be scaled-up to obtain large amounts of highly purified, concentrated eGFP with a purity >99% after a crystal wash step and resolubilization. The proposed chromatography-free purification procedure gives reason to expect significant reductions of costs and required process time compared to conventional preparative chromatography.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biotechnology - Volume 194, 20 January 2015, Pages 84-90
Journal: Journal of Biotechnology - Volume 194, 20 January 2015, Pages 84-90
نویسندگان
Dariusch Hekmat, Dominik Maslak, Matthias Freiherr von Roman, Peter Breitschwerdt, Christoph Ströhle, Alexander Vogt, Sonja Berensmeier, Dirk Weuster-Botz,