Rapid fabrication of functionalized plates for peptides, glycopeptides and protein purification and mass spectrometry analysis

Abstract

A rapid and simple approach for fabricating a disposable functionalized membrane on matrix-assisted laser desorption ionization (MALDI) targets, glass, or plastic substrates, without using complex mechanical protocols or chemical reactions, was developed for sample enrichment and mass spectrometry analysis. By coating functionalized-silica particles on a polydimethylsiloxane (PDMS)-coated plate, these particles can form a monolayer of materials on the PDMS membrane for sample handling without peeling off. An octadecyl(C18)-functionalized plate was fabricated by coating porous C18-silica particles on a PDMS-coated plate. The C18 particle-coated PDMS plate (CP plate) has better sensitivity than C18 tips and magnetic nanoparticles, along with a higher sample recovery (64.3 ± 4.9%) compared to the C18 tip method, when analyzing trace amounts of 5 fm BSA digest samples. The CP plate shows significantly higher urea/SDS removal efficiency on the cell lysate proteome compared to C18 tips. The capacity of the C18 spot (∼2.8 mm in diameter) on the CP plate was ∼10 μg of BSA digests. A hydrophilic particle-coated PDMS plate was also fabricated and successfully used for glycopeptide enrichment and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis.

Urine proteome analysis by C18 plate–matrix-assisted laser desorption/ionization time-of-flight mass spectrometry allows noninvasive differential diagnosis and prediction of diabetic nephropathy

“…However, SELDI-TOF MS is limited by its high-cost protein purification chips and specified TOF MS. In order to provide a low-cost and universal protein purification chip, we have developed a C 18 plate that allows for rapid sample desalting and protein purification processes [ 11 ].…”

A Rapid Approach for Fabricating Boronic Acid-Functionalized Plates for On-Probe Detection of Glycoprotein and Glycopeptide

“…Recently, we have made a plate coated with hydrophilic interaction chromatography (HILIC) beads, which successfully enrich 10 HRP glycopeptides in a complex peptides samples. 9) However, the HILIC plate has a poor ability in the puri cation of glycoproteins, which may be attributed to the similar hydrophilicity of glycoprotein and proteins when they are dissolved in water solution.…”

“…In our previous study, 9) we reported that by simply adding silicate particle suspension solution on PDMS, a monolayer of the particles can be immobilized on PDMS lm without peeling o a er water/organic solvent (ACN or IPA) ushing or sonication due to covalent bonding happen on the contact interface of PDMS and silica particles. In this study, we synthesized boronic acid particles and coated these particles on the PDMS plate.…”

“…[9][10][11] Brie y, an aliquot (3-5 µL) of the PDMS mixture (prepolymer : curing agent, 10 : 1 in volume ratio) was smeared on a glass or a stainless steel MALDI plate and was atten by a roller (part No. 165-1279; Bio-Rad).…”

“…Recently, we have developed a rapid approach for fabricating functionalized plates through bonding silica-based functionalized particles on polydimethylsiloxane (PDMS) membrane. 9) Herein, a novel approach for on-plate selective enrichment and puri cation of glycoproteins/glycopeptides is proposed based on the new boronic-funtionalized MALDI plate.…”