期刊
JOURNAL OF SEPARATION SCIENCE
卷 39, 期 23, 页码 4653-4659出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.201600931
关键词
Capillary electrophoresis; Monoclonal anti-c-Myc antibodies; Peptides; Quantum dots; Self-assembly
资金
- National High Technology Research and Development Program of China (863 Program) [2014AA020521]
- National Natural Science Foundation of China [21602020]
- Project of Jiangsu Province Industry-University-Research joint innovation fund [BY2016029-22]
- International Scientific Cooperation Project of Changzhou Scientific Bureau [CZ20160015]
- Scientific research project of Jiangsu Traditional Chinese Medicine Bureau [YB2015104]
- Advanced Catalysis and Green Manufacturing Collaborative Innovation Center of Changzhou University
- University Natural Science Research Program of Jiangsu Province [15KJB360001]
- 333 Project of Jiangsu Province and the Open Fund of the Key Laboratory of Synthetic Biology, Chinese Academy of Sciences, China [SYN201603]
Herein, we report an assay for detecting the binding of a multivalent peptide and antibody within a capillary with the use of fluorescence coupled capillary electrophoresis. Quantum dots and a c-Myc tag containing peptide EQKLISEEDLG(4)H(6) were injected sequentially and formed a multivalent quantum dot-EQKLISEEDLG(4)H(6) assembly within the capillary. The efficiency of the quantum dot-peptide self-assembly was affected by the peptide/quantum dotmolar ratio, sampling time, and interval time. Finally, the binding of the monoclonal anti-c-Myc antibody and the multivalent quantum dot-EQKLISEEDLG(4)H(6) ligand was studied using an in-capillary assay. The microscopic dissociation constant for the self-assembly of monoclonal anti-c-Myc antibody and quantum dot-EQKLISEEDLG(4)H(6) was determined to be 14.1 mu M with a stoichiometry of the peptide-antibody complex of 1.7 determined after fitting this to the Hill equation. This method can be further extended to detect a wide range of biomolecule-biomolecule binding interactions.
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