کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1200336 1493579 2014 11 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Adenovirus purification by two-column, size-exclusion, simulated countercurrent chromatography
ترجمه فارسی عنوان
تصفیه آدنویروس با استفاده از دو ستون، حذف اندازه، کروماتوگرافی مقابله با جریان همزمان
کلمات کلیدی
شبیه سازی تخت در حال حرکت، دو ستون کروماتوگرافی، تصفیه ویروس، اندازه کروماتوگرافی حذف، پردازش پایین آدنویروس
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی


• Ad5 was purified by size-exclusion chromatography (SEC) in a 2-column open-loop SMB system.
• Clearance of impurities, namely DNA and host cell protein (HCP), were experimentally assessed.
• The pilot-scale run yielded a virus recovery of 86% and clearances of 90% and 89% for DNA and HCP.
• The virus yield was increased from 57% for the batch system to 86% for the 2-column SMB system.
• The productivity was increased by 6-fold with the two-column SMB process.

Adenovirus serotype 5 (Ad5) was successfully separated by size-exclusion chromatography (SEC) using a simple, yet efficient, two-column, quasi-continuous, simulated moving-bed process operated in an open-loop configuration. The operating cycle is divided into two identical half-cycles, each of them consisting of the following sequence of sub-steps: (i) elution of the upstream column and direction of the effluent of the downstream column to waste; (ii) elution of the upstream column and redirection of its effluent to waste while the downstream column is fed with the clarified bioreaction bulk and its effluent collected as purified product; (iii) operation of the system as in step (i) but collecting the effluent of the downstream column as product; (iv) elution of the upstream column and direction of its effluent to waste while the flow through the downstream column is temporarily halted. Clearance of impurities, namely DNA and host cell protein (HCP), were experimentally assessed. The pilot-scale run yielded a virus recovery of 86%, and a clearance of 90% and 89% for DNA and HCP, respectively, without any fine tunning of the predetermined operating parameters. These figures compare very favorably against single-column batch chromatography for the same volume of size-exclusion resin. However, and most importantly, the virus yield was increased from 57% for the batch system to 86% for the two-column SEC process because of internal recycling of the mixed fractions of contaminated Ad5, even though the two-column process was operated strictly in an open-loop configuration. And last, but not least, the productivity was increased by 6-fold with the two-column process. In conclusion, the main drawbacks of size-exclusion chromatography, namely low productivity and low product titer, were overcome to a considerable extent by an innovative two-column configuration that keeps the mixed fractions inside the system at all times.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1347, 20 June 2014, Pages 111–121
نویسندگان
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