Article ID Journal Published Year Pages File Type
1214843 Journal of Chromatography B 2016 6 Pages PDF
Abstract

•High sensitive and selective phosphopeptides enrichment by titanium(IV) cross-linked chitosan composite (Ti-CTS).•Natural chitosan is applied for IMAC supporting material.•Phosphopeptides in standard proteins (casein variants) and two real samples (non-fat milk and serum) were well enriched by Ti-CTS.

Natural chitosan was applied as supporting material for Ti(IV) based immobilized metal ion affinity chromatographic (IMAC) material (Ti-CTS). Compared with other polymer based IMAC, Ti-CTS can save the cockamamie synthesis procedures and be easy to obtain. The morphology, surface area, pore volume and elemental composition of Ti-CTS were revealed by scanning electron microscopy (SEM), Brunauer–Emmett–Teller (BET) method and X-ray photoelectron spectroscopy (XPS). Tryptic digest products from several standard proteins and two real samples (non-fat milk and serum) were enriched using Ti-CTS to demonstrate the efficiency of this method. The results showed that this composite enables high sensitive and selective phosphopeptide enrichment from casein variants, non-fat milk and human serum. Furthermore, multi-phosphorylated peptides with three serine phospholated sites (S*S*S*) demonstrated high affinity to Ti-CTS. Hence, this method had great potential for future studies of complex phosphoproteomes and especially multi-phosphorylated peptides.

Related Topics
Physical Sciences and Engineering Chemistry Analytical Chemistry
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