The use of liquid phase deposition prepared phosphonate grafted silica nanoparticle‐deposited capillaries in the enrichment of phosphopeptides

Abstract

In our current work, we describe how open tubular-immobilized metal-ion affinity chromatography (OT-IMAC) capillary columns connected to a solid phase microextraction (in-tube SPME) device can be used for the enrichment of phosphopeptides. A phosphonate modified silica nanoparticle (NP)-deposited capillary was prepared by liquid phase deposition (LPD), and used for the immobilization of Fe(3+), Zr(4+) or Ti(4+). The enrichment capacities of three different OT-IMAC capillary columns were compared by using tryptically digested alpha-casein as sample. The improved extraction efficiency in our technique was demonstrated by comparing to a directly modified capillary, and a comparison of phosphopeptide extraction from simple and complex samples was tested for both modes. Our results show that the NP-IMAC-Zr(4+) capillary column can be used to selectively isolate phosphopeptides from real samples, and can enrich for beta-casein phosphopeptides from concentrations as low as 1.7x10(-9) M.

Immobilized metal affinity chromatography using open tubular capillary for phosphoprotein analysis: Comparison between polymer brush coating and surface functionalization

“…Similar approaches based on open tubular IMAC-Fe 3+ columns allowed the identification of one phosphorylated protein expressed in E. coli [27]. Another study employing Zr 4+ or Ti 4+ ions created by liquid phase deposition on phosphonategrafted silica nanoparticle-deposited capillaries shown promising results regarding phosphopeptide enrichment [28]. A recent study proposed the same strategy for phosphopeptide enrichment starting by Surface Initiated Atom Transfer Radical Polymerization (SI-ATRP) of glycidyle methacrylate (GMA) followed by opening the epoxide with N,N,N ,N ,N -pentamethyldiethylenetriamine, a reaction of the terminal amine with phosphorus oxychloride (POCl 3 ) and the immobilization of zirconium (IV) [29].…”

Tunable thick porous silica coating fabricated by multilayer-by-multilayer bonding of silica nanoparticles for open-tubular capillary chromatographic separation

“…Moreover, their applications have been intensely proposed in several analytical chemistry fields. Previously, porous silica layer was prepared by dynamic coating [42], static coating [43][44][45], liquid phase deposition [30], or sol-gel method [46]. However, the layer thickness, obtained by all of these coating procedures, was either too thin or uncontrollable.…”

“…However, although these types of nanoparticles have been used successfully for chromatographic separation, in most cases, columns were coated with only a single layer. Some of the columns were coated with a few layers of nanoparticles [28,30,34,[37][38][39][40][41][42]. However, the thickness of the coating was still very thin.…”

“…Compared to other materials, nanoparticles have the merits of large surface area and ease of preparation. Therefore, nanoparticles of silica [30], gold [26,[31][32][33][34][35], titanium dioxide [36][37][38], and grapheme oxide nanosheets [39][40][41] have been immobilized onto the inner surface of capillary column. However, although these types of nanoparticles have been used successfully for chromatographic separation, in most cases, columns were coated with only a single layer.…”

Ti (IV) attached‐phosphonic acid functionalized capillary monolith as a stationary phase for in‐syringe‐type fast and robust enrichment of phosphopeptides

“…A series of pioneering studies on the development of immobilized metal affinity chromatography techniques for the enrichment of phosphopeptides were performed (Dong, Zhou, Wu, Ye, & Zou, ; Feng et al, ; Wu et al, ; Wu et al, ; Zhou et al, ; Zhou et al, ). Zirconium phosphonate‐modified porous silicon was first evaluated for specific capture of phosphopeptides (Zhou et al, ).…”