Quantitative ligand immobilization using alginate hydrogel formed in a capillary: Application for online affinity concentration

Yudai Fukushima, Toyohiro Naito, Kenji Sueyoshi, Takuya Kubo, Fumihiko Kitagawa, Koji Otsuka

研究成果: Contribution to journalArticle査読

3 被引用数 (Scopus)

抄録

To simplify a quantitative immobilization procedure of ligands with maintaining their activities, we developed an automated preparation method using an alginate hydrogel partially formed in a capillary. After a sodium alginate solution containing a ligand was injected into the capillary, a background solution containing Ca2+ was then introduced into the sodium alginate solution zone by applying an appropriate voltage for the hydrogelation, resulting in encapsulation of the ligand by the formed alginate hydrogel. According to the estimated binding capacity for biotin by the encapsulated avidin, the injected avidin was immobilized quantitatively by the formed hydrogel with keeping its affinity. When avidin (3.5-35.2 ng) was immobilized by the proposed method, the immobilization efficiency was estimated to be almost 100%. Furthermore, by using the prepared capillary, biotinylated fluorescein was specifically trapped and separated due to the affinity of the encapsulated avidin. By the change of pH of the background solution, the concentrated analytes could be easily eluted, resulting in 90% recovery with high reproducibility by using 1.18 fmol biotinylated sample.

本文言語英語
ページ(範囲)5977-5982
ページ数6
ジャーナルAnalytical Chemistry
86
12
DOI
出版ステータス出版済み - 6 17 2014
外部発表はい

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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