Highly specific revelation of rat serum glycopeptidome by boronic acid-functionalized mesoporous silica

• Published in: Analytica chimica acta 2012-11, Vol.753, p.64-72
• Main Authors: Liu, Liting, Zhang, Ying, Zhang, Lei, Yan, Guoquan, Yao, Jun, Yang, Pengyuan, Lu, Haojie
• Format: Article
• Language: English
• Subjects: Adsorption; Analytical chemistry; Animals; Binding; Biological and medical sciences; Boronic Acids - chemistry; Chemistry; Chromatography, High Pressure Liquid; Diverse techniques; Enrichment; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Glycopeptides; Glycopeptides - blood; Glycosylation; Mass spectrometry; Materials selection; Mesoporous materials; Molecular and cellular biology; Porosity; Proteomics; Rats; Selectivity; Serum glycopeptidome; Serums; Silicon dioxide; Silicon Dioxide - chemistry; Size exclusion; Spectrometric and optical methods; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; State of the art
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2012.10.002

A highly ordered boronic acid-functionalized mesoporous silica was synthesized and applied for the revelation of rat serum glycopeptidome for the first time. [Display omitted] ► A highly ordered boronic acid-functionalized mesoporous silica was synthesized. ► The as-prepared material possessed both glycopeptide-suitable pore size and glycopeptide-specific selectivity. ► The as-prepared material showed highly efficient ability for enrichment of endogenous glycopeptides from serum. ► Rat serum glycopeptidome was reveled for the first time. Although the specific profiling of endogenous glycopeptides in serum is highly inclined towards the discovery of disease biomarkers, studies on the endogenous glycopeptides (glycopeptidome) have never been conducted because of several factors. These factors include the high dynamic range of serum proteins, the inadequacy of traditional sample preparation techniques in proteomics for low-molecular-weight (LMW) proteins, and the relatively low abundances of glycopeptides. Boronic acid-functionalized mesoporous silica was synthesized in this study to overcome the limitations of the state-of-the-art methods for glycopeptidome research. The boronic acid-functionalized mesoporous silica exhibited excellent selectivity by analyzing glycopeptides in the mixture of glycopeptides/non-glycopeptides at molar ratio of 1:100, extreme sensitivity (the limit of detection was at the fmol level), good binding capacity (40mgg−1), as well as the high post-enrichment recovery of glycopeptides (up to 88.10%). The as-prepared material possessing both glycopeptide-suitable pore size and glycopeptide-specific selectivity has shown special capability for enriching the endogenous glycopeptides. Fifteen unique glycosylation sites mapped to 15 different endogenous glycopeptides were identified in rat serum. The established protocol revealed for the first time the rat serum glycopeptidome.