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Analyst. 2018 Jul 7;143(13):3141-3147. doi: 10.1039/c8an00714d. Epub 2018 Jun 7.

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

Author information

1
Molecular Recognition Research Center, Korea Institute of Science and Technology (KIST), Seoul 02792, Korea. mjkang1@kist.re.kr and Division of Bio-Medical Science and Technology (Biological Chemistry), University of Science and Technology (UST), Daejeon 34113, Korea.
2
Department of Materials Science and Engineering, Hallym University, Chuncheon-si, Korea and Integrative Materials Research Institute, Hallym University, Chuncheon-si, Korea.
3
Molecular Recognition Research Center, Korea Institute of Science and Technology (KIST), Seoul 02792, Korea. mjkang1@kist.re.kr.

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.

PMID:
29877533
DOI:
10.1039/c8an00714d
[Indexed for MEDLINE]

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