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Licensed Unlicensed Requires Authentication Published by De Gruyter November 9, 2021

High-resolution capillary electrophoresis for the determination of carbamylated albumin

Julien Favresse ORCID logo and Joris Delanghe ORCID logo

Abstract

Objectives

Carbamylation is a non-enzymatic post-translational reaction of a primary amino group of a protein with isocyanate. The albumin carbamylation is a negative prognostic factor in chronic kidney disease (CKD) patients and induce charge difference implying an observed shift in electrophoretic mobility that can be measured through a symmetry factor (SF).

Methods

The Helena V8 and the Sebia Capillarys 2 systems were used for all experiments. The effect of in vitro carbamylation on the SF by spiking increasing concentrations of potassium isocyanate (KCNO) in serum of three healthy volunteers was investigated. Theoretical plate numbers (N) as a surrogate of separation efficiency were also calculated and correlations between SF and renal function biomarkers were performed on 284 patients.

Results

A dose-dependent impact of KCNO on the SF was observed for both methods with the Helena V8 being more sensitive. The mean N was significantly higher on the Helena V8 as compared to the Sebia Capillarys 2 (2,972 vs. 444.1, p<0.0001). The SF correlated significantly with eGFR (r=0.50, p<0.0001), creatinine (r=−0.31, p<0.0001) and urea (r=−0.34, p<0.0001) on the Helena V8. On the Sebia Capillarys 2, a significant correlation was only observed with eGFR (r=0.17, p=0.004). A better discrimination between CKD stages was also observed using the Helena V8.

Conclusions

Thanks to a higher mean N, the Helena V8 might offer new possibilities, including detection of carbamylated albumin through SF calculation. Further studies are still needed to confirm the interest of using this type of assays in clinical routine.


Corresponding author: Julien Favresse, Department of Laboratory Medicine, Clinique St-Luc Bouge, 8 Rue Saint-Luc, 5000 Namur, Belgium; and Department of Pharmacy, Namur Research Institute for LIfes Sciences, University of Namur, Namur, Belgium, Phone: +32 81 20 91 44, E-mail:

Acknowledgments

The authors thanks Mr Thierry De Bruyne for performing the Adobe InDesign analysis and Mr Guillaume Sondag for performing the spiking experiment.

  1. Research funding: None declared.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: Authors state no conflict of interest.

  4. Informed consent: Informed consent was obtained from the three healthy volunteers included in this study.

  5. Ethical approval: The local Institutional Review Board approved the study exempt from review.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2021-0926).


Received: 2021-08-19
Accepted: 2021-10-28
Published Online: 2021-11-09
Published in Print: 2022-01-27

© 2021 Walter de Gruyter GmbH, Berlin/Boston

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