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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 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 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 immunoassay were 0.1 µg/mL and 0.3 µg/mL, respectively. The calibration curves for the immunocomplex had the 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 was ranged between 89-103%. 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 a useful for quantitative immunoassay development.

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Supplementary files

Publication details

The article was received on 18 Apr 2018, accepted on 15 May 2018 and first published on 16 May 2018


Article type: Paper
DOI: 10.1039/C8AN00714D
Citation: Analyst, 2018, Accepted Manuscript
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    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, Accepted Manuscript , DOI: 10.1039/C8AN00714D

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