Klukova Ludmila, Bertok Tomas, Petrikova Miroslava, Sediva Alena, Mislovicova Danica, Katrlik Jaroslav, Vikartovska Alica, Filip Jaroslav, Kasak Peter, Andicsová-Eckstein Anita, Mosnáček Jaroslav, Lukáč Jozef, Rovenský Jozef, Imrich Richard, Tkac Jan
Department of Glycobiotechnology, Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, Bratislava 845 38, Slovakia.
Center for Advanced Materials, Qatar University, P.O. Box 2713, Doha, Qatar; Department for Biomaterial Research, Polymer Institute, Slovak Academy of Sciences, Dúbravská cesta 9, Bratislava 845 41, Slovakia.
Anal Chim Acta. 2015 Jan 1;853:555-562. doi: 10.1016/j.aca.2014.10.029. Epub 2014 Oct 27.
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.
系统性硬化症（SSc）是一种严重影响患者生活质量的自身免疫性疾病。该疾病的异质性也意味着疾病生物标志物的识别及后续验证颇具挑战性。目前尚未有一个经过充分验证的单一生物标志物可用于诊断、预后评估、疾病活动监测以及治疗反应评估。本研究的主要目的是通过使用识别唾液酸的凝集素黑接骨木凝集素（SNA）对血清糖谱样本进行检测，将一种替代检测方案应用于基于免疫分析的该疾病研究中。据我们所知，这是第一项描述基于凝集素直接对系统性硬化症患者血清样本进行糖谱分析的研究。本文从生物分析的角度比较了三种基于凝集素的血清样本糖谱分析方法，包括传统的类似酶联免疫吸附测定的凝集素法（ELLA）、新型荧光凝集素微阵列和超灵敏阻抗凝集素生物传感器。当将健康人的血清与系统性硬化症患者的血清进行比较时，所有这三种生物分析方法获得的结果均一致显示糖蛋白上唾液酸水平存在差异。因此，分析人血清中的唾液酸含量可能对未来系统性硬化症的检测具有诊断价值，但还需要进一步开展工作，例如通过对富含个体诊断抗体的人血清部分进行糖谱分析来提高检测的选择性。
