Combination of click chemistry and Schiff base reaction: Post‐synthesis of covalent organic frameworks as an immobilized metal ion affinity chromatography platform for efficient capture of global phosphopeptides in serum with chronic obstructive pulmonary disease

Reasonable design and construction of functionalized materials are of great importance for the enrichment of global phosphopeptides. In this work, Ti 4+ functionalized hydrophilic covalent organic frameworks by introducing glutathione (GSH) and 2,3,4‐trihydroxy benzaldehyde (THBA) via click chemistry and Schiff base reaction (COF‐V@GSH‐THBA‐Ti 4+ ) was constructed and applied for selective enrichment of phosphopeptides in serum. Benefit from the high surface area, excellent hydrophilicity as well as regular mesoporous structure, COF‐V@GSH‐THBA‐Ti 4+ displayed high selectivity (molar ratio of 2000:1), low limit of detection (0.5 fmol), high load capacity (100.0 mg/g) and excellent size‐exclusion effect (1:10000) for enrichment of phosphopeptides. For actual bio‐sample analysis, 15 phosphopeptides assigned to 10 phosphoproteins with 16 phosphorylated sites and 33 phosphopeptides assigned to 25 phosphoproteins with 34 phosphorylated sites were detected from the serum of patients with chronic obstructive pulmonary disease (COPD), and normal controls. Biological processes and molecular functions analysis further disclosed the difference of serums with phosphoproteomics between COPD and normal controls.