Single-Step Purification of Monomeric l-Selectin via Aptamer Affinity Chromatography
Abstractl-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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Kuehne, C.; Wedepohl, S.; Dernedde, J. Single-Step Purification of Monomeric l-Selectin via Aptamer Affinity Chromatography. Sensors 2017, 17, 226.
Kuehne C, Wedepohl S, Dernedde J. Single-Step Purification of Monomeric l-Selectin via Aptamer Affinity Chromatography. Sensors. 2017; 17(2):226.Chicago/Turabian Style
Kuehne, Christian; Wedepohl, Stefanie; Dernedde, Jens. 2017. "Single-Step Purification of Monomeric l-Selectin via Aptamer Affinity Chromatography." Sensors 17, no. 2: 226.
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