Abstract
Improvement in high-throughput MALDI-TOF MS analysis requires practical and efficient sample preparation protocols for high acquisition rates. The use of hexagonal mesoporous silica (HMS) sorbents in combination with MALDI-TOF MS was explored as a versatile tool for peptidomic profiling of clinical specimens difficult to process, but considered important sources of disease biomarkers: synovial fluid and sputum. A rapid and efficient procedure, based on dispersive solid-phase extraction of peptides using commercially available wormhole mesostructured HMS, was tested for: a) pre-concentration of standard peptides in serially diluted solution up to the sub-nanomolar range; b) peptidome profiling of sputum and synovial fluid. The use of HMS, as dispersed sponges, significantly amplified the peptidic repertoire of sputum and synovial fluid by excluding from the adsorptive process large size proteins, which mask and/or suppress peptidome signals. The protocol proposed, as dispersive solid phase extraction, ensures good analytical performances. Moreover, it is economical and rapid, as it avoids the use of less reproducible and prolonged sample preparation procedures, such as the use of ultrafiltration filter devices. These findings may contribute to defining a high-throughput screening MS-based platform for monitoring key peptidic features of difficult to analyse bodily fluids in a clinical setting.
Highlights
Peptidomics is a promising “omics” approach for the study and the qualitative–quantitative analysis of endogenous peptides in biological samples
A strategy to decrease the limit of detection of extremely diluted analytes is to concentrate them, as it happens in the case of ELISA, in which the analytes are concentrated by mean of antigen-antibody interaction
We have described the use of HMS for pre-concentrating, desalting and peptide enrichment from both diluted and complex mixtures, such as synovial fluid (SF) and sputum
Summary
Peptidomics is a promising “omics” approach for the study and the qualitative–quantitative analysis of endogenous peptides in biological samples. In concomitance to well-known magnetic beads [7,8], or conventional SPE devices [9], innovative platforms and mesoporous materials for selective capture of peptides, phosphopeptides and proteins from bodily fluids and tissues, have been recently proved to be very successful [10,11,12] Given their highly ordered mesostructures, the large surface area of the pores, which accounts for up to 95% of the total surface of the material, mesoporous materials can directly interact and selectively capture biologically significant subproteomes from complex clinical matrices [13]. As a part of an ongoing project aimed to screen the ability of mesoporous materials for selective isolation of peptides from clinical bio-fluids prior to MS analysis, wormhole mesostructured (hexagonal mesoporous silica) HMS, previously explored by our group for nasal fluid peptidome enrichment [16], were further investigated in this technical study for synovial fluid (SF). A smart procedure was developed with the aim to provide a high-throughput MALDI-TOF MS-based platform for monitoring key peptidic-patterns useful for clinical biomarker investigations, after validation of its robustness by analysing several replicates of sputum and SF, subject of future work
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
