کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2021092 | 1069227 | 2010 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Recombinant production of bioactive human TNF-α by SUMO-fusion system - High yields from shake-flask culture
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موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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![عکس صفحه اول مقاله: Recombinant production of bioactive human TNF-α by SUMO-fusion system - High yields from shake-flask culture Recombinant production of bioactive human TNF-α by SUMO-fusion system - High yields from shake-flask culture](/preview/png/2021092.png)
چکیده انگلیسی
We have therefore established a protocol for the rapid high-level synthesis of recombinant human TNF-α in Escherichia coli shake-flask cultures and the subsequent purification of the mature protein. Using the advantages of SUMO-fusion technology we were able to produce protein with an authentic N-terminus in high yield. Two immobilized metal ion-affinity chromatography steps with a protease cleavage step in between and subsequent size-exclusion chromatography were utilized to purify the protein. The protein was obtained from the last chromatography step as a trimer, while purity was at least 96% as estimated by SDS-PAGE. The identity of the protein was confirmed by MALDI-TOF mass spectrometry. Recombinant mature TNF-α was correctly folded as assessed by CD spectroscopy and its biological activity was confirmed by an L929 cell assay.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 72, Issue 2, August 2010, Pages 238-243
Journal: Protein Expression and Purification - Volume 72, Issue 2, August 2010, Pages 238-243
نویسندگان
Andreas Hoffmann, Mathias Q. Müller, Manja Gloser, Andrea Sinz, Rainer Rudolph, Sven Pfeifer,