AKIYAMA Yoshikatsu
   Department   Research Institutes and Facilities, Research Institutes and Facilities
   Position   Assistant Professor
Article types Original article
Language English
Peer review Peer reviewed
Title Affinity Chromatography with Collapsibly Tethered Ligands
Journal Formal name:Analytical Chemistry
Abbreviation:Anal. Chem.
ISSN code:0003-2700/1520-6882
Domestic / ForeginForegin
Publisher American Chemical Society
Volume, Issue, Page 75(7),pp.1658-1663
Author and coauthor YAMANAKA Hidenori, YOSHIZAKO Kimihiro, AKIYAMA Yoshikatsu, SOTA Hiroyuki, HASEGAWA Yukio, SHINOHARA Yasuro, KIKUCHI Akihiko, OKANO Teruo
Publication date 2003/03/01
Summary We introduce a novel affinity chromatography mode in which affinity ligands are secured to the media surface via collapsible tethers. In traditional affinity chromatography, the immobilized ligands act passively, and their local concentration is static. In collapsibly tethered affinity chromatography, the ligand can move dynamically in response to external stimuli, a design that enables marked changes in both the local concentration of the ligand and its surrounding environment without exchange of solvent. Using the thermoresponsive polymer poly(N-isopropylacrylamide) (PIPAAm) as a scaffold for ligand and hapten attachment, we were able to achieve controlled mobility and microenvironment alteration of the affinity ligand Ricinus communis agglutinin (RCA120). The glycoprotein target, asialotransferrin, was loaded onto a column in which PIPAAm was partially substituted with both RCA120 and lactose. At 5 °C, the column retained the glycoprotein, but released most (95%) of the asialotransferrin upon warming to 30 °C. This temperature-induced elution was much greater than can be explained by temperature dependency of sugar recognition by RCA120. The simplest explanation is that upon thermally induced dehydration and collapse of the PIPAAm chains, coimmobilized RCA120 ligand and lactose hapten are brought into closer proximity to each other, enabling immobilized lactose to displace affinity-bound asislotransferrin from the immobilized RCA120 lectin.
DOI https://doi.org/10.1021/ac0263768
URL for researchmap https://researchmap.jp/y_akiyama