One-Step Preparation of Zwitterionic-Rich Hydrophilic Hydrothermal Carbonaceous Materials for Enrichment of N-Glycopeptides

Abstract

Two kinds of novel zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) materials decorated with phosphorylcholine or sulfobetaine type zwitterionic groups were facilely synthesized employing hydrothermal carbonization (HTC) reaction, in which sustainable carbohydrate (glucose) could be converted into functionalized carbonaceous materials with strong hydrophilicity using a one-step and environmentally mild process. The properties of obtained materials were characterized by helium ion microscopy (HIM), N2 adsorption/desorption measurement, static water contact angle measurement, elemental analysis, Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), and ζ-potential. As a result, 26 N-glycopeptides with S/N above 20 could be detected from 150 fmol IgG digests after hydrophilic interaction chromatography (HILIC) enrichment by using as-synthesized material HTC-Glc-10%MPC. The limit of detection in matrix assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF MS) analysis was as low as 5 fmol and enrichment selectivity reached 100:1. Furthermore, 175 N-glycosylation sites in 285 N-glycopeptides from 92 glycoproteins were identified from 2 μL of human serum. This work provided a new, simple, and green way to synthesize an N-glycopeptide enrichment material for liquid chromatography–mass spectrometry/mass spectrometry (LC-MS/MS) analysis of glycoproteomes.

A Pragmatic Guide to Enrichment Strategies for Mass Spectrometry–Based Glycoproteomics

“…A common property achieved by these materials is hydrophilicity, although the avenues to achieve such properties differ. Materials include cobalt sulfide nanoleaves ( 242 ), covalent/metal organic framework approaches ( 181 , 243 , 244 , 245 , 246 ), carbon microspheres ( 247 ), and polymeric monoliths functionalized with piperazine ( 248 ), among others. As described above, some of these materials can have bifunctional properties to enable HILIC-based glycopeptide enrichment and metal ion–based phosphopeptide enrichment ( 181 , 182 , 249 ).…”

Bi‐amino acid functionalized biomimetic honeycomb chitosan membrane as a multifunctional hydrophilic probe for specific capture of N‐linked glycopeptides in nasopharyngeal carcinoma's disease patient's serum

“…So far, boric acid chemistry [16], hydrazine chemistry, lectin affinity method [17], and hydrophilic interaction chromatography (HILIC) [18,19] have become the main enrichment methods for glycopeptides. Among them, HILIC has been extensively applied in the separation and enrichment of glycopeptides [20,21], and many HILICbased materials, including metal organic frameworks (MOFs) [22] and graphene oxide [23], have been explored to capture glycopeptides [24,25]. Although a certain degree of performance has been achieved, HILIC-based nanomaterials also have the limitation of insufficient hydrophilicity.…”

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

“…At present, the main enrichment methods that have been developed and used include lectin affinity method [14], boronic acid method [15,16], hydrazine chemical method [17], hydrophilic interaction method, and so on [18]. Among them, it is worth mentioning that because the hydrophilic interaction method has significant advantages such as low bias to different glycopeptides, better compatibility, high throughput, simplicity, and efficiency, it is widely used in the enrichment and separation of glycopeptides [19][20][21][22][23][24]. The principle of hydrophilic chromatography takes advantage of the difference in hydrophilicity of proteins before and after glycosylation.…”