Issue 13, 2018

Capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based immunoassay for quantifying antibodies against cyclic citrullinated peptides

Abstract

We developed an assay for diagnosing rheumatoid arthritis (RA) by capillary electrophoresis with the laser-induced fluorescence detection (CE-LIF) of antibodies against cyclic citrullinated peptides (CCPs) using fluorescein isothiocyanate-labeled F-CCP11A and F-CCP. The CCPs were incubated overnight with a purified antibody against CCP and then injected into the CE-LIF system to estimate complex concentration. The immunocomplexes and free peptides were separated within 5 min; the LOD and LOQ values of the immunoassay are 0.1 μg mL−1 and 0.3 μg mL−1, respectively. The calibration curves for the immunocomplex had a coefficient of determination of 0.98. The reproducibility of the CE-LIF based immunoassay when using F-CCP11A and F-CCP was less than 5% and the accuracy ranged between 89 and 103%. The applicability of the assay was evaluated using serum samples from RA patients and healthy control subjects. We found that the sensitivity and specificity for RA diagnosis were 100% and 83.3%, respectively, for F-CCP11A and 100% and 90%, respectively, for F-CCP. These results demonstrate that the CE-LIF method can be useful for quantitative immunoassay development.

Graphical abstract: Capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based immunoassay for quantifying antibodies against cyclic citrullinated peptides

Supplementary files

Article information

Article type
Paper
Submitted
18 Apr 2018
Accepted
15 May 2018
First published
16 May 2018

Analyst, 2018,143, 3141-3147

Capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based immunoassay for quantifying antibodies against cyclic citrullinated peptides

B. T. Nguyen, M. Park, Y. S. Yoo and M. Kang, Analyst, 2018, 143, 3141 DOI: 10.1039/C8AN00714D

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