title: In-capillary self-assembly and proteolytic cleavage of polyhistidine peptide capped quantum dots.
abstract: A new method using fluorescence coupled capillary electrophoresis (CE-FL) for monitoring self-assembly and proteolytic cleavage of hexahistidine peptide capped quantum dots (QDs) inside a capillary has been developed in this report. QDs and the ATTO 590-labeled hexahistidine peptide (H6-ATTO) were injected into a capillary, sequentially. Their self-assembly inside the capillary was driven by a metal-affinity force which yielded a new fluorescence signal due to Förster resonance energy transfer (FRET). The highly efficient separation of fluorescent complexes and the FRET process were analyzed using CE-FL. The self-assembly of QDs and biomolecules was found to effectively take place inside the capillary. The kinetics of the assembly was monitored by CE-FL, and the approach was extended to the study of proteolytic cleavage of surface conjugated peptides. Being the first in-depth analysis of in-capillary nanoparticle-biomolecule assembly, the novel approach reported here provides inspiration to the development of QD-based FRET probes for biomedical applications.
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, 213164, China.
authors.forename: Jianhao
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, 213164, China.
authors.forename: Jingyan
authors.initials: J
authors.lastname: Li
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, 213164, China.
authors.forename: Jinchen
authors.initials: J
authors.lastname: Li
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, 213164, China.
authors.forename: Feifei
authors.initials: F
authors.lastname: Liu
authors.affiliation: Changzhou Qianhong Bio-pharma Co. Ltd, Changzhou 213164, Jiangsu, People's Republic of China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Changzhou Qianhong Bio-pharma Co. Ltd, Changzhou 213164, Jiangsu, People's Republic of China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Yao
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, 213164, China.
authors.forename: Cheli
authors.initials: C
authors.lastname: Wang
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, 213164, China. Electronic address: linqiupjj@gmail.com.
authors.forename: Lin
authors.initials: L
authors.lastname: Qiu
authors.affiliation: School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, 213164, China; State Key Laboratory of Pharmaceutical Biotechnology, Nanjing, Jiangsu, People's Republic of China. Electronic address: pengju.jiang@gmail.com.
authors.forename: Pengju
authors.initials: P
authors.lastname: Jiang
mesh_terms: D019075:Electrophoresis, Capillary; D005453:Fluorescence; D006639:Histidine / Q000737:chemistry*; D059748:Proteolysis; D045663:Quantum Dots
chemical_list: C033223:polyhistidine; D006639:Histidine
keywords: Fluorescence coupled capillary electrophoresis; Förster resonance energy transfer; Peptide; Quantum dots; Thrombin