کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5136733 | 1494025 | 2016 | 5 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Rapid N-glycan release from glycoproteins using immobilized PNGase F microcolumns
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کلمات کلیدی
IgGBFSGSTAPTSPNGase FSPRiCGE-LIFRibonuclease BRNase B8-Aminopyrene-1,3,6-trisulfonic acid - 8-آمینوپیرن-1،3،6-تری سولفونیک اسیدN-glycosylation - N-گلیکوزیلتCapillary electrophoresis - الکتروفورزمویرگیimmunoglobulin G - ایمونوگلوبولین GEnzyme immobilization - بی حرکت سازی آنزیمglutathione-S-transferase - گلوتاتیون S-ترانسفراز
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
N-glycosylation profiling of glycoprotein biotherapeutics is an essential step in each phase of product development in the biopharmaceutical industry. For example, during clone selection, hundreds of clones should be analyzed quickly from limited amounts of samples. On the other hand, identification of disease related glycosylation alterations can serve as early indicators (glycobiomarkers) for various pathological conditions in the biomedical field. Therefore, there is a growing demand for rapid and easy to automate sample preparation methods for N-glycosylation analysis. In this paper, we report on the design and implementation of immobilized recombinant glutathione-S-transferase (GST) tagged PNGase F enzyme microcolumns for rapid and efficient removal of N-linked carbohydrates from glycoproteins. Digestion speed and efficiency were compared to conventional in-solution based protocols. The use of PNGase F functionalized microcolumns resulted in efficient N-glycan removal in 10Â min from all major N-linked glycoprotein types of: (i) neutral (IgG), (ii) highly sialylated (fetuin), and (iii) high mannose (ribonuclease B) carbohydrate containing glycoprotein standards. The approach can be readily applied to automated sample preparation systems, such as liquid handling robots.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography B - Volume 1032, 1 October 2016, Pages 139-143
Journal: Journal of Chromatography B - Volume 1032, 1 October 2016, Pages 139-143
نویسندگان
Marton Szigeti, Judit Bondar, Douglas Gjerde, Zsolt Keresztessy, Akos Szekrenyes, Andras Guttman,