کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
7610316 | 1493495 | 2016 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Advance chromatin extraction improves capture performance of protein A affinity chromatography
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موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Practical effects of advance chromatin removal on performance of protein A affinity chromatography were evaluated using a caprylic acid-allantoin-based extraction method. Lacking this treatment, the practice of increasing loading residence time to increase capacity was shown to increase host protein contamination of the eluted IgG. Advance chromatin extraction suspended that compromise. Protein A ligand leakage from columns loaded with chromatin-extracted harvest was half the level observed on protein A columns loaded with non-extracted harvest. Columns loaded with chromatin-extracted harvest were cleaned more effectively by 50-100Â mM NaOH than columns loaded with non-extracted harvest that were cleaned with 250-500Â mM NaOH. Two protein A media with IgG capacities in excess of 50Â g/L were loaded with chromatin-extracted harvest, washed with 2.0Â M NaCl before elution, and the eluted IgG fraction titrated to pH 5.5 before microfiltration. Host protein contamination in the filtrate was reduced to <1Â ppm, DNA to <1Â ppb, protein A leakage to 0.5Â ppm, and aggregates to 1.0%. Caprylic acid and allantoin were both reduced below 5Â ppm. Step recovery of IgG was 99.4%. Addition of a single polishing step reduced residual protein A beneath the level of detection and aggregates to <0.1%. Overall process recovery including chromatin extraction was 90%.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1431, 29 January 2016, Pages 1-7
Journal: Journal of Chromatography A - Volume 1431, 29 January 2016, Pages 1-7
نویسندگان
Rui Nian, Wei Zhang, Lihan Tan, Jeremy Lee, Xeuzhi Bi, Yuansheng Yang, Hui Theng Gan, Pete Gagnon,