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. 2019 Dec 22;21(1):101.
doi: 10.3390/ijms21010101.

Modification of Luffa Sponge for Enrichment of Phosphopeptides

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Free PMC article

Modification of Luffa Sponge for Enrichment of Phosphopeptides

Lili Dai et al. Int J Mol Sci. .
Free PMC article

Abstract

The enrichment technique is crucial to the comprehensive analysis of protein phosphorylation. In this work, a facile, green and efficient synthetic method was set up for quaternization of luffa sponge. The resultant luffa sponge showed strong anion-exchange characteristics and a high adsorption ability for phosphate ions. Along with the unique physical properties, e.g., tenacity and porous texture, quaternized luffa sponge was demonstrated to be a well-suited solid-phase extraction (SPE) material. The quaternized luffa sponge-based SPE method was simple, cost-effective and convenient in operation, and was successfully applied to the capture of phosphopeptides from protein digests. The enrichment approach exhibited exceptionally high selectivity, sensitivity and strong anti-interference ability. Four phosphopeptides were still detected by using the digest mixture of β-casein and bovine serum albumin with a molar ratio of 1:100. 21 phosphopeptides were identified from the tryptic digest of non-fat milk.

Keywords: Matrix-assisted laser desorption ionization; luffa sponge; mass spectrometry; phosphopeptide; solid-phase extraction.

Conflict of interest statement

The authors declare that they have no conflicts of interest.

Figures

Figure 1
Figure 1
SEM images of (a,b) untreated luffa sponge, (c,d) modified luffa sponge QA, and (e,f) modified luffa sponge QB.
Figure 2
Figure 2
XPS spectra of N 1s of (a) untreated luffa sponge, (b) modified luffa sponge QB, and (c) modified luffa sponge QA.
Figure 3
Figure 3
Ion exchange chromatograms of anions (a) before and after adsorption by (b) untreated luffa sponge, (c) QA, and (d) QB.
Figure 4
Figure 4
Schematic diagram of the procedure for enrichment of phosphopeptides with QB. Loading solution: 30 mM acetic acid-ammonium acetate in 20% acetonitrile, pH 4.0; washing solution: deionized water; elution solution: 5% trifluoroacetic acid.
Figure 5
Figure 5
MALDI mass spectra of tryptic digests of α-casein (3 pmol) by direct analysis (a) or after enrichment using QB (b), and β-casein (3 pmol) by direct analysis (c) or after enrichment using QB (d).
Figure 6
Figure 6
MALDI mass spectra of (a) tryptic digest of non-fat milk without any enrichment, (b) with QB enrichment, and (c) with TiO2 enrichment.

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