کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10550043 | 967012 | 2005 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Downstream processing of MDCK cell-derived equine influenza virus
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کلمات کلیدی
kDaFCSNaHCO3kiloDaltonMDCKMultiangle Laser Light ScatteringPBSDNA - DNA یا اسید دزوکسی ریبونوکلئیکSDS–PAGE - SDS-PAGEInfluenza - آنفلوانزا EDTA - اتیلن دی آمین تترا استیک اسید ethylene diamine tetra-acetic acid - اتیلن دیامین تترا استیک اسیدdeoxyribonucleic acid - اسید deoxyribonucleicUltrafiltration - اولترافیلتراسیونsodium bicarbonate - سدیم بیکربناتfoetal calf serum - سرم گوساله جنینDownstream processing - فرایند فرآیند پایینphosphate buffer saline - فسفات بافر شورgel filtration - فیلتر کردن ژلMALS - مالشMadin Darby canine kidney - مدین داربی کلیه کلیویneuraminidase - نورآمینیدازhemagglutinin - هماگلوتینینMolecular weight - وزن مولکولیBEI - وقتیVirus - ویروسChromatography - کروماتوگرافی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
A microcarrier-based process was used to produce equine influenza virus (A/Equi 2 (H3N8), Newmarket 1/93) in Madin Darby Canine kidney (MDCK) cells. The virus was purified in a sequence of downstream processing steps comprising of depth filtration, inactivation, ultrafiltration (UF) and gel filtration. In the ultrafiltration step, the hemagglutinin (HA) was recovered to 100%. A high increase of neuraminidase (NA) activity indicated the removal of some inhibitory compounds during this step. At the same time, the level of contaminating proteins and DNA was reduced by more than 88%. In the subsequent size exclusion chromatography (Sepharose CL 2B), the recovery of HA and NA in the “virus peak” was 37.8 and 59.8%, respectively compared to the concentrated feed material. Inconsistencies in the overall mass balance for HA and NA (70.0 and 69.2%) during gel filtration indicated non-specific interactions of the inactivated virus to the gel matrix which is supported by a HA recovery of about 50% in shake flask experiments performed as a control. Overall 35.8% of HA and 291.6% of NA were recovered. More than 95.7% of the host cell proteins and 98.7% of the host cell DNA were removed during downstream processing.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography B - Volume 823, Issue 2, 5 September 2005, Pages 75-81
Journal: Journal of Chromatography B - Volume 823, Issue 2, 5 September 2005, Pages 75-81
نویسندگان
Deba Prasad Nayak, Sylvia Lehmann, Udo Reichl,