Facile preparation of titanium functionalized cross-linked chitosan polymer for phosphoproteome analysis in serum

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 1247; p. 124347
• Main Authors: Sheng, Xiuqin, Guo, Yimin, Ding, Chuan-Fan, Yan, Yinghua
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.10.2024
• Subjects: Alzheimer Disease - blood; Blood Proteins - analysis; Blood Proteins - chemistry; blood serum; chitosan; Chitosan - chemistry; chromatography; crosslinking; Humans; Immobilized metal affinity chromatography; MALDI-TOF MS; Phosphopeptides; Phosphopeptides - blood; Phosphopeptides - chemistry; Phosphoproteins - blood; Phosphoproteins - chemistry; phosphoproteome; polymers; Proteome - analysis; Proteome - chemistry; proteomics; Proteomics - methods; titanium; Titanium - chemistry; MALDI-TOF MS; Phosphopeptides; Immobilized metal affinity chromatography
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2024.124347

•Titanium-rich hydrophilic polymeric material was prepared via a facile method.•The Schiff base reaction was introduced to generate abundant amines and cis-dihydroxyl groups.•The polymer exhibited excellent sensitivity, selectivity and loading capacity for phosphopeptides enrichment. Efficient phosphopeptide enrichment is extremely important for proteomics research. In this work, chitosan (CTs), 2,3-dihydroxyterephthalaldehyde (2,3-DHA), and carbohydrazide (CHZ) are polymerized to generate the polymer (DHA-CTs-CHZ), and then the polymer (DHA-CTs-CHZ) is coupled with a significant number of titanium ions to enrich phosphopeptides. The material exhibits high selectivity (5000:1 M ratio of BSA to β-casein), sensitivity (0.001 fmol/μL), loading (83.3 μg/mg), recovery (98.6 ± 1.2 %), and effective size exclusion for phosphopeptide enrichment. In addition, 46 phosphopeptides and 31 phosphorylated sites associated with 27 phosphorylated proteins were successfully captured from the serum of normal subjects, while 47 phosphopeptides and 35 phosphorylated sites associated with 30 phosphorylated proteins were successfully captured from Alzheimer’s disease (AD) patients’ serum.