アキヤマ ヨシカツ   AKIYAMA Yoshikatsu
  秋山 義勝
   所属   研究施設 研究施設
   職種   講師
論文種別 原著
言語種別 英語
査読の有無 査読あり
表題 Affinity Chromatography with Collapsibly Tethered Ligands
掲載誌名 正式名:Analytical Chemistry
略  称:Anal. Chem.
ISSNコード:0003-2700/1520-6882
掲載区分国外
出版社 American Chemical Society
巻・号・頁 75(7),pp.1658-1663
著者・共著者 YAMANAKA Hidenori, YOSHIZAKO Kimihiro, AKIYAMA Yoshikatsu, SOTA Hiroyuki, HASEGAWA Yukio, SHINOHARA Yasuro, KIKUCHI Akihiko, OKANO Teruo
発行年月 2003/03/01
概要 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
researchmap用URL https://researchmap.jp/y_akiyama