Glycoprofiling as a novel tool in serological assays of systemic sclerosis: A comparative study with three bioanalytical methods

• Assays of systemic sclerosis (SSc) in human serum by three bioanalytical methods.
• Lectin-based glycoprofiling using electrochemical impedance spectroscopy (EIS), microarrays and ELISA-like approach.
• Ultrasensitive glycoprofiling by EIS with LOD down to 10 aM.
• Increased level of sialic acid-containing glycoproteins in control sample compared to SSc samples.
• The first direct glycoprofiling assays of SSc in human serum.

Systemic sclerosis (SSc) is an autoimmune disease seriously affecting patient’s quality of life. The heterogeneity of the disease also means that identification and subsequent validation of biomarkers of the disease is quite challenging. A fully validated single biomarker for diagnosis, prognosis, disease activity and assessment of response to therapy is not yet available. The main aim of this study was to apply an alternative assay protocol to the immunoassay-based analysis of this disease by employment of sialic acid recognizing lectin Sambucus nigra agglutinin (SNA) to glycoprofile serum samples. To our best knowledge this is the first study describing direct lectin-based glycoprofiling of serum SSc samples. Three different analytical methods for glycoprofiling of serum samples relying on application of lectins are compared here from a bioanalytical point of view including traditional ELISA-like lectin-based method (ELLA), novel fluorescent lectin microarrays and ultrasensitive impedimetric lectin biosensors. Results obtained by all three bioanalytical methods consistently showed differences in the level of sialic acid present on glycoproteins, when serum from healthy people was compared to the one from patients having SSc. Thus, analysis of sialic acid content in human serum could be of a diagnostic value for future detection of SSc, but further work is needed to enhance selectivity of assays for example by glycoprofiling of a fraction of human serum enriched in antibodies for individual diagnostics.

Glycoprofiling of human serum by: (A) lectin-based electrochemical impedance spectroscopy, (B) microarray with fluorescently labeled lectin and (C) enzyme-linked lectin assay (ELLA) with enzyme-labeled lectin.Figure optionsDownload as PowerPoint slide