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Single-Step Purification of Monomeric l-Selectin via Aptamer Affinity Chromatography
Kuehne, Christian ;  Wedepohl, Stefanie ;  Dernedde, Jens

HaupttitelSingle-Step Purification of Monomeric l-Selectin via Aptamer Affinity Chromatography
AutorKuehne, Christian
AutorWedepohl, Stefanie
AutorDernedde, Jens
Seitenzahl7 S.
Freie Schlagwörterl-selectin; aptamer; DNA; recombinant protein; purification; affinity; SPR
DDC610 Medizin und Gesundheit
Auch erschienen inSensors. - 17 (2017), 2, Artikel Nr. 226
Zusammenfassungl-selectin is a transmembrane receptor expressed on the surface of white blood cells and responsible for the tethering of leukocytes to vascular endothelial cells. This initial intercellular contact is the first step of the complex leukocyte adhesion cascade that ultimately permits extravasation of leukocytes into the surrounding tissue in case of inflammation. Here we show the binding of a soluble histidine tagged l-selectin to a recently described shortened variant of an l-selectin specific DNA aptamer with surface plasmon resonance. The high specificity of this aptamer in combination with its high binding affinity of ~12 nM, allows for a single-step protein purification from cell culture supernatants. In comparison to the well-established Ni-NTA based technology, aptamer affinity chromatography (AAC) was easier to establish, resulted in a 3.6-fold higher protein yield, and increased protein purity. Moreover, due to target specificity, the DNA aptamer facilitated binding studies directly from cell culture supernatant, a helpful characteristic to quickly monitor successful expression of biological active l-selectin. View Full-Text
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Fachbereich/EinrichtungFB Biologie, Chemie, PharmazieMedizinische Fakultät Charité - Universitätsmedizin Berlin
Erscheinungsjahr2017
Dokumententyp/-SammlungenWissenschaftlicher Artikel
SpracheEnglisch
RechteCreative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Anmerkungen des AutorsDer Artikel wurde in einer reinen Open-Access-Zeitschrift publiziert.
Erstellt am05.05.2017 - 12:01:05
Letzte Änderung05.05.2017 - 12:02:33
 
Statische URLhttp://edocs.fu-berlin.de/docs/receive/FUDOCS_document_000000026960
DOI10.3390/s17020226
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