title: Simultaneous detection of assembly and disassembly of multivalent HA tag and anti-HA antibody in single in-capillary assay.
abstract: Herein, we have developed an in-capillary assay for simultaneous detection of the assembly and disassembly of the multivalent HA tag peptide and antibody. HA tag with hexahistidine at C terminus (YPYDVPDYAG4 H6 , termed YPYDH6 ) was conjugated with quantum dots (QDs) by metal-affinity force to form a multivalent HA tag (QD-YPYDH6 ). QD-YPYDH6 and monoclonal anti-HA antibody (anti-HA) were sequentially injected into the capillary. They were mixed and assembled inside the capillary. The reaction products were online discriminated and detected by fluorescence coupled capillary electrophoresis (CE-FL). For the in-capillary assay, the binding efficiency of the multivalent HA tag and antibody on was influenced by the molar ratio and injection time. Such novel assay could even give out the self-assembly kinetic constant of QDs and YPYDH6 as KD of 34.1 μM with n (binding cooperativeness) of 2.2 by Hill equation. More importantly, the simultaneous detection of the assembly and imidazole (Im) induced disassembly of the QD-YPYDH6 -anti-HA complex was achieved in a single in-capillary assay. Our study demonstrated a new method for the online detection of antigen-antibody interactions.
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Jiangsu, P. R. China.
authors.forename: Jianhao
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Petrochemical Engineering, Changzhou University, Jiangsu, P. R. China.
authors.forename: Yuqin
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Institute of Pharmaceutical and Environmental Engineering, Changzhou Vocational Institute of Engineering, Jiangsu, P. R. China.
authors.forename: Haifang
authors.initials: H
authors.lastname: Qin
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Jiangsu, P. R. China.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Jiangsu, P. R. China.
authors.forename: Shumin
authors.initials: S
authors.lastname: Ding
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Jiangsu, P. R. China.
authors.forename: Yiwan
authors.initials: Y
authors.lastname: Teng
authors.affiliation: Changzhou Meisheng Biomaterials Co., Ltd, Jiangsu, P. R. China.
authors.forename: Junling
authors.initials: J
authors.lastname: Ji
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Jiangsu, P. R. China.|Changzhou Meisheng Biomaterials Co., Ltd, Jiangsu, P. R. China.
authors.forename: Lin
authors.initials: L
authors.lastname: Qiu
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Jiangsu, P. R. China.
authors.forename: Pengju
authors.initials: P
authors.lastname: Jiang
mesh_terms: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005453:Fluorescence; D006639:Histidine / Q000276:immunology; D009842:Oligopeptides / Q000276:immunology; D045663:Quantum Dots / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; C471213:His-His-His-His-His-His; D009842:Oligopeptides; D006639:Histidine
keywords: Anti-HA; Antigen-antibody interaction; CE-FL; Quantum dots; YPYDVPDYAG4H6