کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
7614843 | 1493976 | 2018 | 25 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Asymmetrical flow field-flow fractionation in purification of an enveloped bacteriophage Ï6
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کلمات کلیدی
DLSradius of hydrationVLPAF4Quaternary aminePESCIMTCAMWCOBSA - BSAbovine serum albumin - آلبومین سرم گاوtrichloroacetic acid - اسید ترشکلراکتیکSDS-PAGE - الکتروفورز ژل پلی آکریل آمیدSodium dodecyl sulfate polyacrylamide gel electrophoresis - الکتروفورز ژل پلی اتیل آمید سدیم دودسیل سولفاتField flow fractionation - تجزیه میدان جریان میدانVirus purification - تصفیه ویروسAsymmetrical flow field-flow fractionation - تقسیم بندی میدان جریان جریان نامتقارنSingle stranded - تک رشتهchannel flow - جریان کانالDouble stranded - دو رشتهConvective interaction media - رسانه متقابل همجوشیRegenerated cellulose - سلولز بازسازی شدهRadius of gyration - شعاع نفوذCross-flow - عبور جریانMALS - مالشplaque forming unit - واحد پالک تشکیل شده استVirus like particle - ویروس مانند ذرهDynamic Light Scattering - پراکندگی نور دینامیکیMulti angle light scattering - پراکندگی نور چند زاویهpfu - پفوpolyethersulfone - پلی اترسولفونpolyethylene glycol - پلی اتیلن گلیکولPEG - پلیاتیلن گلیکول Macromolecular complex - پیچیدگی مولکولیmolecular weight cut-off - کاهش وزن مولکولی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Basic and applied virus research requires specimens that are purified to high homogeneity. Thus, there is much interest in the efficient production and purification of viruses and their subassemblies. Advances in the production steps have shifted the bottle neck of the process to the purification. Nonetheless, the development of purification techniques for different viruses is challenging due to the complex biological nature of the infected cell cultures as well as the biophysical and -chemical differences in the virus particles. We used bacteriophage Ï6 as a model virus in our attempts to provide a new purification method for enveloped viruses. We compared asymmetrical flow field-flow fractionation (AF4)-based virus purification method to the well-established ultracentrifugation-based purification of Ï6. In addition, binding of Ï6 virions to monolithic anion exchange columns was tested to evaluate their applicability in concentrating the AF4 purified specimens. Our results show that AF4 enables one-hour purification of infectious enveloped viruses with specific infectivity of ~1â¯Ãâ¯1013â¯PFU/mg of protein and ~65-95% yields. Obtained purity was comparable with that obtained using ultracentrifugation, but the yields from AF4 purification were 2-3-fold higher. Importantly, high quality virus preparations could be obtained directly from crude cell lysates. Furthermore, when used in combination with in-line light scattering detectors, AF4 purification could be coupled to simultaneous quality control of obtained virus specimen.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography B - Volume 1095, 15 September 2018, Pages 251-257
Journal: Journal of Chromatography B - Volume 1095, 15 September 2018, Pages 251-257
نویسندگان
Mirka Lampi, Hanna M. Oksanen, Florian Meier, Evelin Moldenhauer, Minna M. Poranen, Dennis H. Bamford, Katri Eskelin,