کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1225654 | 968240 | 2010 | 9 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: A rapid screening method for cell lines producing singly-tagged recombinant proteins using the “TARGET tag” system A rapid screening method for cell lines producing singly-tagged recombinant proteins using the “TARGET tag” system](/preview/png/1225654.png)
Recombinant production of extracellular glycoproteins in stable mammalian cell lines is an ideal technique for obtaining a large quantity of high-quality proteins. In most cases, however, current methodologies do not allow for sufficiently rapid cell line development and protein purification. Here, we describe a 21-residue peptide tag (designated as TARGET tag) and its use for rapid stable cell line development and purification. The ability of the anti-tag antibody P20.1 to withstand repetitive regeneration cycles has enabled the development of a sensitive surface plasmon resonance-based screening format that requires only 20 µl of cell culture supernatants. Direct and semi-quantitative screening at the 96-well culture scale eliminated the need for a second screening, re-cloning, or sorting, thereby minimizing culture pre-production time. Using this system, “high producer” cell lines were established in less than a month, and milligram quantities of target proteins could be purified with a standardized protocol.
Journal: Journal of Proteomics - Volume 73, Issue 9, 5 August 2010, Pages 1777–1785