کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6132953 | 1593448 | 2016 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Standardized large-scale H-1PV production process with efficient quality and quantity monitoring
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کلمات کلیدی
CPECsClHCPMOI - MEq-PCR - Q-PCRCytopathic effect - اثر سیتوپاتیکHAU - اینCharacterization - تعیین مشخصاتLarge-scale production - تولید در مقیاس بزرگRoom temperature - دمای اتاقquantitative real time PCR - زمان واقعی PCR کمیRefraction index - شاخص شکستگیElectron microscopy - میکروسکوپ الکترونیendotoxin unit - واحد اندوتوکسینinfectious unit - واحد عفونیhemagglutination unit - واحد هموگلوبینplaque forming unit - واحد پالک تشکیل شده استPurification - پاکسازیhost cell protein - پروتئین سلولی میزبانpfu - پفوmultiplicity of infection - چندین عفونتcesium chloride - کلرید سزیم
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ویروس شناسی
پیش نمایش صفحه اول مقاله
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چکیده انگلیسی
The promising anticancer properties of rodent protoparvoviruses, notably H-1PV, have led to their clinical testing. This makes it necessary to produce highly pure, well-characterized virus batches in sufficient quantity. The present work focused on developing standardized production, purification, and characterization procedures as a basis for exploiting H-1PV both preclinically and in clinical trials for anticancer virotherapy. Two infection and two virus purification strategies were tested and the resulting virus preparations compared for their purity and full-, infectious-, and empty-particle contents. The adopted production process, which involves culturing and infecting NB-324K cells in 10-layer CellSTACK® chambers (1 Ã 103 infectious units per infected cell), is simple, scalable, and reproducible. Downstream processing to eliminate contaminating DNA and protein includes DNAse treatment, filtration, and two Iodixanol density-gradient centrifugations, the first gradient being a step gradient and the second, either a step (1 Ã 1010 PFU/ml) or a continuous gradient (3 Ã 1011 PFU/ml). A procedure was also developed for obtaining infectious particle-free preparations of empty virions for research purposes: cesium chloride density gradient centrifugation followed by UV irradiation (1 Ã 1014 physical particles/ml). For quick, sensitive determination of physical particles (and hence, particle-to-infectivity ratios), a “Capsid-ELISA” was developed, based on a novel monoclonal antibody that specifically targets assembled capsids.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 229, March 2016, Pages 48-59
Journal: Journal of Virological Methods - Volume 229, March 2016, Pages 48-59
نویسندگان
Barbara Leuchs, Mandy Roscher, Marcus Müller, Kathrin Kürschner, Jean Rommelaere,