Mesopore-assisted Profiling of Human Synovial Fluid Mariaimmacolata Preianòa, Stella Frascàa, Domenico Scopellitia, Antonio Coriglianob, Luca Gallellia, Olimpio Galassob, Giorgio Gasparinib, Rocco Savinoa and Rosa Terraccianoa* a) Dept. of Health Sciences, University of Catanzaro Magna Græcia, Catanzaro, Italy; b) Dept. of Medical and Surgical Sciences, University of Catanzaro Magna Græcia, Catanzaro, Italy Progress in the field of nanostructured materials has provided innovative devices, particularly those based on mesoporous silica (MPS), which have proven to be successful for protein fractionation. Our group has explored a new application of MPS by developing a strategy based on MPS in combination with MALDI-TOF MS for profiling low molecular weight human bodily fluids peptides [1]. Based on a molecular cut-off mechanism, we used MPS as sponges to “capture” peptides present in human bodily fluids. By the mean of controllable pore size and surfaces properties, MPS were used to harvest peptides from human body fluids, excluding large size and high abundant proteins from adsorptive process [2]. We are now revisiting our strategy based on MPS for rapidly profiling synovial fluid (SF) peptidome. SF is an highly viscous fluid with lubricant properties which contains a large number of proteins originating from synovial tissue, cartilage, and serum [3]. SF also contains large sized carbohydrate polymers of hyaluronic acid, which seriously hinder the MALDI-MS detection of peptides in unprocessed samples. In particular, here we describe the protocol which we have developed for SF peptidome profiling by differently functionalized MPS particles for SF peptidome enrichment. The assessment of SF pre-processing with or without hyaluronidase before MPS-MALDI-TOF MS analysis will also be treated with special focus on peptide recovery and analytical reproducibility. Finally, a comparison with different commercially available SPE kits and MSP performances will be shown. * Corresponding author: Rosa Terracciano, Prof. University Magna Graecia Catanzaro, Europa Avenue, 88100 Catanzaro, Italy. Tel.: +39 0961 369 4085; fax: +39 0961 369 4090. E-mail address: terracciano@unicz.it (R. Terracciano) References: [1] Savino R, Terracciano R. Mesopore-assisted profiling strategies in clinical proteomics for drug/target discovery. Drug Discov Today. 2012;17(3-4):143-52. [2] Terracciano R, Pasqua L, Casadonte F, Frascà S, Preianò M, Falcone D, Savino R. Derivatized mesoporous silica beads for MALDI-TOF MS profiling of human plasma and urine. Bioconjug Chem. 2009;20(5):913-23. [3] Hu S, Loo JA, Wong DT. Human body fluid proteome analysis. Proteomics. 2006;6(23):6326-53.

Mesopore-assisted Profiling of Human Synovial Fluid

Gallelli L;Galasso O;
2012-01-01

Abstract

Mesopore-assisted Profiling of Human Synovial Fluid Mariaimmacolata Preianòa, Stella Frascàa, Domenico Scopellitia, Antonio Coriglianob, Luca Gallellia, Olimpio Galassob, Giorgio Gasparinib, Rocco Savinoa and Rosa Terraccianoa* a) Dept. of Health Sciences, University of Catanzaro Magna Græcia, Catanzaro, Italy; b) Dept. of Medical and Surgical Sciences, University of Catanzaro Magna Græcia, Catanzaro, Italy Progress in the field of nanostructured materials has provided innovative devices, particularly those based on mesoporous silica (MPS), which have proven to be successful for protein fractionation. Our group has explored a new application of MPS by developing a strategy based on MPS in combination with MALDI-TOF MS for profiling low molecular weight human bodily fluids peptides [1]. Based on a molecular cut-off mechanism, we used MPS as sponges to “capture” peptides present in human bodily fluids. By the mean of controllable pore size and surfaces properties, MPS were used to harvest peptides from human body fluids, excluding large size and high abundant proteins from adsorptive process [2]. We are now revisiting our strategy based on MPS for rapidly profiling synovial fluid (SF) peptidome. SF is an highly viscous fluid with lubricant properties which contains a large number of proteins originating from synovial tissue, cartilage, and serum [3]. SF also contains large sized carbohydrate polymers of hyaluronic acid, which seriously hinder the MALDI-MS detection of peptides in unprocessed samples. In particular, here we describe the protocol which we have developed for SF peptidome profiling by differently functionalized MPS particles for SF peptidome enrichment. The assessment of SF pre-processing with or without hyaluronidase before MPS-MALDI-TOF MS analysis will also be treated with special focus on peptide recovery and analytical reproducibility. Finally, a comparison with different commercially available SPE kits and MSP performances will be shown. * Corresponding author: Rosa Terracciano, Prof. University Magna Graecia Catanzaro, Europa Avenue, 88100 Catanzaro, Italy. Tel.: +39 0961 369 4085; fax: +39 0961 369 4090. E-mail address: terracciano@unicz.it (R. Terracciano) References: [1] Savino R, Terracciano R. Mesopore-assisted profiling strategies in clinical proteomics for drug/target discovery. Drug Discov Today. 2012;17(3-4):143-52. [2] Terracciano R, Pasqua L, Casadonte F, Frascà S, Preianò M, Falcone D, Savino R. Derivatized mesoporous silica beads for MALDI-TOF MS profiling of human plasma and urine. Bioconjug Chem. 2009;20(5):913-23. [3] Hu S, Loo JA, Wong DT. Human body fluid proteome analysis. Proteomics. 2006;6(23):6326-53.
2012
978-88-903318-5-5
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11386/4861222
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