title: In-capillary probing QDs and HAT tag self-assembly and displacement using Förster resonance energy transfer.
abstract: HAT tag, a natural His affinity tag, is one of the most popular fusion tags. However, HAT tag containing three positive charges limited its self-assembly with quantum dots (QDs). Herein, ATTO 590-labeled HAT peptide was synthesized to self-assemble with QDs inside the capillary. QDs and ATTO 590-labeled HAT peptide were sequentially injected into the capillary and probed by fluorescence-coupled CE (CE-FL), showing an obvious Förster resonance energy transfer signal. Online self-assembly, separation, and detection were achieved within 10 min. CE-FL further revealed that imidazole and H6G6peptide could partially outcompete with HAT tag on the QDs surface inside the capillary. The displacement intermediates were separated clearly using CE-FL. Our study demonstrates the power of online CE-FL in analyzing the binding interaction between ligands containing positive charges and QDs.
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Jianhao
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Jinchen
authors.initials: J
authors.lastname: Li
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Chencheng
authors.initials: C
authors.lastname: Zhang
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Jie
authors.initials: J
authors.lastname: Fan
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Yaqin
authors.initials: Y
authors.lastname: Gu
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Feifei
authors.initials: F
authors.lastname: Liu
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Cheli
authors.initials: C
authors.lastname: Wang
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Bingyu
authors.initials: B
authors.lastname: Dong
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Lin
authors.initials: L
authors.lastname: Qiu
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Pengju
authors.initials: P
authors.lastname: Jiang
keywords: Fluorescence-coupled CE; Förster resonance energy transfer; Natural His affinity tag; Quantum dots; Self-assembly