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Int J Biol Macromol. 2018 Jul 1;113:900-906. doi: 10.1016/j.ijbiomac.2018.03.032. Epub 2018 Mar 7.

A novel monitoring approach of antibody-peptide binding using "bending" capillary electrophoresis.

Author information

1
School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou 213164, Jiangsu, People's Republic of China.
2
Changzhou Second People's Hospital Affiliated to Nanjing Medical University, Changzhou 213164, People's Republic of China.
3
Department of Biomedical Science, Mercer University School of Medicine, Savannah Campus, Hoskins Building, Room #2209, 4700 Waters Ave, Savannah, GA, USA, 31404.
4
School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou 213164, Jiangsu, People's Republic of China; Department of Biomedical Science, Mercer University School of Medicine, Savannah Campus, Hoskins Building, Room #2209, 4700 Waters Ave, Savannah, GA, USA, 31404. Electronic address: LI_J@mercer.edu.
5
School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou 213164, Jiangsu, People's Republic of China. Electronic address: linqiupjj@gmail.com.
6
State Key Laboratory of Radiation Medicine and Protection, School of Radiation Medicine and Protection, Collaborative Innovation Center of Radiological Medicine of Jiangsu Higher Education Institutions, Soochow University, Suzhou 215123, People's Republic of China. Electronic address: lyq@suda.edu.cn.
7
School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou 213164, Jiangsu, People's Republic of China. Electronic address: pengju.jiang@gmail.com.

Abstract

Recently, the in-capillary electrophoresis assay has been applied to variety kinds of analyses owing to its multiple functional integrating features, including mixing of samples, reaction process of the mixtures, and the separation and detection in one capillary system. However, the micro-reactor still has its limitations to the currently available applications, especially the mixing step of the samples inside the capillary could not be well controlled automatically or manually. Herein, we have developed a novel capillary electrophoresis assay for the detection of antibody-peptide binding inside a bending capillary. Its efficacy was monitored using an anti-FLAG M2 antibody and its ligand conjugated with FAM dye (FAM-DYKD). The antibody and the peptide were mixed inside the bending capillary with sequential injections. It was found that the numbers of semi-circle on the capillary interfered by the antibody and peptide binding dynamic. Additionally, an online competition assay was performed, which further validated the efficacy of the bending capillary device on monitoring the dynamic binding between the antigen and antibody. In summary, our data suggests that the novel assay is a practical approach in monitoring the antibody-antigen complex formation at a nano-scale. It could be applied to detect any biomolecule-biomolecule interaction as a general strategy.

KEYWORDS:

Antibody; Bending capillary; Capillary electrophoresis; Fluorescence

PMID:
29524489
DOI:
10.1016/j.ijbiomac.2018.03.032
[Indexed for MEDLINE]

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