Hydrophilic Nanocomposite Functionalized by Carrageenan for the Specific Enrichment of Glycopeptides

Abstract

A hydrophilic nanocomposite was synthesized by an easy route to improve glycopeptides enrichment efficiency. This new composite, prepared with a method based on electrostatic interaction, was demonstrated to be efficient for immobilization of carrageenan on graphene oxide/poly-(ethylenimine) support (denoted as GO−PEI−Carr). Carrageenan, which has a large number of hydroxyl groups and is fully negatively charged, is a new modified phase of hydrophilic materials in glycoproteomics. The introduction of carrageenan provided the composite not only a perfect surface charge but also a greater ability to enrich glycosylated peptides. Thirty-four glycopeptides from human serum immunoglobulin G (IgG) tryptic digests were obviously observed with greatly improved signal-to-noise (S/N) ratio. A good selectivity was still kept even when the molar ratio of IgG and bovine serum albumin (BSA) tryptic digest mixtures reached to 1:500. Meanwhile, 76 glycopeptides derived from 56 glycoproteins with 83 N-glycosylation sites were identified from human serum and 149 glycopeptides derived from 129 glycoproteins with 157 N-glycosylation sites were identified from mouse liver tissues, which showed the ability to enrich glycopeptides from complex biological samples. In addition, GO−PEI−Carr exhibited a unique repeatability and stability even after enrichment of glycopeptides for 20 times. It also performed a higher sensitivity (1 fmol/μL IgG), a better enrichment capacity (up to ∼300 mg/g), and an ideal enrichment recovery (90.8% and 109.5%) for glycopeptides enrichment, indicating a great potential for the application of glycoproteomic research.

Fabrication of magnetic dual-hydrophilic metal organic framework for highly efficient glycopeptide enrichment

“…The glycopeptide information obtained was searched using MASCOT software, and 101 glycopeptides were identified, corresponding to 48 glycoproteins. These results were better than some reported studies [34,35]. The specific glycosylation information is recorded in ESM Table S6.…”

Selective enrichment of sialylated glycopeptides with mesoporous poly-melamine-formaldehyde (mPMF) material

“…As a choice, significant improvement of SGP selectivity was achieved with multiple-modals material by taking advantage of the synergistic effect of hydrophilicity and electrostatic binding with intrinsic hydrophilic and electrostatic properties of glycopeptides. For example, sepharose [18] and carrageenan [19] conjugating together ionic functional group and carbohydrate-based solid support obtained better selectivity for glycopeptides. Electrostatic repulsion-hydrophilic interaction chromatography (ERLIC) [20] isolation using polyethyleneimine-bonded silica has been extended to comprehensively profile glyco-and phosphoproteomes of mouse brain membrane proteome [21].…”

Hydrophilic carrageenan functionalized magnetic carbon‐based framework linked by silane coupling agent for the enrichment of N‐glycopeptides from human saliva

“…Up to now, Carr has few applications in glycopeptides enrichment, so there is a promising outlook. Chen et al synthesized GO@PEI@Carr, which detected 76 N-glycopeptides from human serum via nano-LC-MS and exhibited good selectivity (1:500) [33]. However, nanomaterial GO@PEI@Carr was obtained by electrostatic interaction, which has poor stability.…”