Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. Previously, we have developed a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans. In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing.

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Journal of Proteome Research
Department of Pediatrics

Bladergroen, M. R., Reiding, K. R., Hipgrave-Ederveen, A., Vreeker, G. C. M., Clerc, F., Holst, S., … Van Der Burgt, Y. E. M. (2015). Automation of high-throughput mass spectrometry-based plasma n-glycome analysis with linkage-specific sialic acid esterification. Journal of Proteome Research, 14(9), 4080–4086. doi:10.1021/acs.jproteome.5b00538