کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
7614910 | 1493979 | 2018 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Broad application and optimization of a single wash-step for integrated endotoxin depletion during protein purification
ترجمه فارسی عنوان
کاربرد گسترده و بهینه سازی مرحله تک شستشو برای تخریب اندوتوکسین یکپارچه در طول پروتئین تصفیه
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کلمات کلیدی
DOECIPIEDMBPSACGSTCEXGMPAExHIC - اینGood Manufacturing Practice - تمرین تولید خوبCleaning in place - تمیز کردن در محلDesign of experiments - طراحی آزمایشاتendotoxin units - واحد اندوتوکسینanion exchange chromatography - کروماتوگرافی تبادل آنیونCation exchange chromatography - کروماتوگرافی تبادل کاتیونیhydrophobic interaction chromatography - کروماتوگرافی تعامل هیدروفوبی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی
Endotoxins contaminate proteins that are produced in E. coli. High levels of endotoxins can influence cellular assays and cause severe adverse effects when administered to humans. Thus, endotoxin removal is important in protein purification for academic research and in GMP manufacturing of biopharmaceuticals. Several methods exist to remove endotoxin, but often require additional downstream-processing steps, decrease protein yield and are costly. These disadvantages can be avoided by using an integrated endotoxin depletion (iED) wash-step that utilizes Triton X-114 (TX114). In this paper, we show that the iED wash-step is broadly applicable in most commonly used chromatographies: it reduces endotoxin by a factor of 103 to 106 during NiNTA-, MBP-, SAC-, GST-, Protein A and CEX-chromatography but not during AEX or HIC-chromatography. We characterized the iED wash-step using Design of Experiments (DoE) and identified optimal experimental conditions for application scenarios that are relevant to academic research or industrial GMP manufacturing. A single iED wash-step with 0.75% (v/v) TX114 added to the feed and wash buffer can reduce endotoxin levels to below 2â¯EU/ml or deplete most endotoxin while keeping the manufacturing costs as low as possible. The comprehensive characterization enables academia and industry to widely adopt the iED wash-step for a routine, efficient and cost-effective depletion of endotoxin during protein purification at any scale.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography B - Volume 1091, 1 August 2018, Pages 101-107
Journal: Journal of Chromatography B - Volume 1091, 1 August 2018, Pages 101-107
نویسندگان
David Koziel, Uwe Michaelis, Tobias Kruse,