title: In-capillary detection of fast antibody-peptide binding using fluorescence coupled capillary electrophoresis.
abstract: Herein, we report a technique for detecting the fast binding of antibody-peptide inside a capillary. Anti-HA was mixed and interacted with FAM-labeled HA tag (FAM-E4 ) inside the capillary. Fluorescence coupled capillary electrophoresis (CE-FL) was employed to measure and record the binding process. The efficiency of the antibody-peptide binding on in-capillary assays was found to be affected by the molar ratio. Furthermore, the stability of anti-HA-FAM-E4 complex was investigated as well. The results indicated that E4 YPYDVPDYA (E4) or TAMRA-E4 YPYDVPDYA (TAMRA-E4) had the same binding priorities with anti-HA. The addition of excess E4 or TAMRA-E4 could lead to partial dissociation of the complex and take a two-step mechanism including dissociation and association. This method can be applied to detect a wide range of biomolecular interactions.
authors.affiliation: School of Petrochemical Engineering, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Yuqin
authors.initials: Y
authors.lastname: Qin
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: Institute of Pharmaceutical and Environmental Engineering, Changzhou Vocational Institute of Engineering, Changzhou, Jiangsu, P. R. China.
authors.forename: Haifang
authors.initials: H
authors.lastname: Qin
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Shumin
authors.initials: S
authors.lastname: Ding
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Yiwan
authors.initials: Y
authors.lastname: Teng
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, P. R. China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Chen
authors.affiliation: School of Petrochemical Engineering, 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: 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: 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: Pengju
authors.initials: P
authors.lastname: Jiang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D001666:Binding Sites, Antibody; D019075:Electrophoresis, Capillary / Q000379:methods*; D005453:Fluorescence; D006825:Hybridomas; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D010455:Peptides
keywords: Anti-HA; Antibody-Peptide Interaction; CE-FL; YPYDVPDYA