2 research outputs found
Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides
The
development of methods to isolate and enrich low-abundance
glycopeptides from biological samples is crucial to glycoproteomics.
Herein, we present an easy and one-step surface modification strategy
to prepare hydrophilic maltose functionalized Fe<sub>3</sub>O<sub>4</sub> nanoparticles (NPs). First, based on the chelation of the
catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative
was assembled on the surface of magnetic Fe<sub>3</sub>O<sub>4</sub> nanoparticles by sonication. Second, the hydrophilic maltose-functionalized
Fe<sub>3</sub>O<sub>4</sub> (Fe<sub>3</sub>O<sub>4</sub>-DA-Maltose)
NPs were obtained via copperÂ(I)-catalyzed azide–alkyne cycloaddition
(click chemistry). The morphology, structure, and composition of Fe<sub>3</sub>O<sub>4</sub>-DA-Maltose NPs were investigated by Fourier
transform infrared spectroscopy (FT-IR), transmission electron microscopy
(TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer
(XPS), and vibrating sample magnetometer (VSM). Meanwhile, hydrophilicity
of the obtained NPs was evaluated by water contact angle measurement.
The hydrophilic Fe<sub>3</sub>O<sub>4</sub>-DA-Maltose NPs were applied
in isolation and enrichment of glycopeptides from horseradish peroxidase
(HRP), immunoglobulin (IgG) digests. The MALDI-TOF mass spectrometric
analysis indicated that the novel NPs exhibited high detection sensitivity
in enrichment from HRP digests at concentration as low as 0.05 ng
μL<sup>–1</sup>, a large binding capacity up to 43 mg
g<sup>–1</sup>, and good recovery for glycopeptides enrichment
(85–110%). Moreover, the Fe<sub>3</sub>O<sub>4</sub>-DA-Maltose
NPs were applied to enrich glycopeptides from human renal mesangial
cells (HRMC) for identification of N-glycosylation sites. Finally,
we identified 115 different N-linked glycopeptides, representing 93
gene products and 124 glycosylation sites in HRMC
Maltose-Functionalized Hydrophilic Magnetic Nanoparticles with Polymer Brushes for Highly Selective Enrichment of N‑Linked Glycopeptides
Efficient enrichment glycoproteins/glycopeptides
from complex biological
solutions are very important in the biomedical sciences, in particular
biomarker research. In this work, the high hydrophilic polyethylenimine
conjugated polymaltose polymer brushes functionalized magnetic Fe<sub>3</sub>O<sub>4</sub> nanoparticles (NPs) denoted as Fe<sub>3</sub>O<sub>4</sub>–PEI–pMaltose were designed and synthesized
via a simple two-step modification. The obtained superhydrophilic
Fe<sub>3</sub>O<sub>4</sub>–PEI–pMaltose NPs displayed
outstanding advantages in the enrichment of N-linked glycopeptides,
including high selectivity (1:100, mass ratios of HRP and bovine serum
albumin (BSA) digest), low detection limit (10 fmol), large binding
capacity (200 mg/g), and high enrichment recovery (above 85%). The
above-mentioned excellent performance of novel Fe<sub>3</sub>O<sub>4</sub>–PEI–pMaltose NPs was attributed to graft of
maltose polymer brushes and efficient assembly strategy. Moreover,
Fe<sub>3</sub>O<sub>4</sub>–PEI–pMaltose NPs were further
utilized to selectively enrich glycopeptides from human renal mesangial
cell (HRMC, 200 μg) tryptic digest, and 449 N-linked glycopeptides,
representing 323 different glycoproteins and 476 glycosylation sites,
were identified. It was expected that the as-synthesized Fe<sub>3</sub>O<sub>4</sub>–PEI–pMaltose NPs, possessing excellent
performance (high binding capacity, good selectivity, low detection
limit, high enrichment recovery, and easy magnetic separation) coupled
to a facile preparation procedure, have a huge potential in N-glycosylation
proteome analysis of complex biological samples