Investigation of multivalent interactions between conjugate of quantum dots with c‐Myc peptide tag and the anti‐c‐Myc antibody by capillary electrophoresis with fluorescence detection |
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Authors: | Jianhao Wang Li Yang Li Liu Hao Wu Jianpeng Wang Lin Qiu |
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Affiliation: | School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, People's Republic of China |
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Abstract: | 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 EQKLISEEDLG4H6 were injected sequentially and formed a multivalent quantum dot‐EQKLISEEDLG4H6 assembly within the capillary. The efficiency of the quantum dot‐peptide self‐assembly was affected by the peptide/quantum dot molar ratio, sampling time, and interval time. Finally, the binding of the monoclonal anti‐c‐Myc antibody and the multivalent quantum dot‐EQKLISEEDLG4H6 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‐EQKLISEEDLG4H6 was determined to be 14.1 μM 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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Keywords: | Capillary electrophoresis Monoclonal anti‐c‐Myc antibodies Peptides Quantum dots Self‐assembly |
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