Cold Plasma-Promoted Glycation of Bovine Caseins: Glycoproteomic Insights into Conformational Relaxation and Epitope Masking.

Cold plasma (CP) pretreatment combined with glycation was applied to modulate bovine casein (CN) allergenicity. Using NanoRPLC-ESI-MS/MS, 62 glycated peptides and 52 modification sites were identified across four CN subtypes, with CP significantly increasing both site numbers and degrees of substitution per site (DSP), especially in αs-CN. Moderate CP exposure (3 min) maximized glycation sensitivity at epitope-rich Lys/Arg residues, reflecting conformational relaxation and enhanced accessibility. CP pretreatment also reshaped Maillard reaction product profiles by promoting early-stage formation while controlling α-dicarbonyl-derived AGEs. Integration of linear B-cell epitope prediction with glycation mapping revealed overlap between high-scoring epitopes and glycated regions, and molecular docking confirmed stable GOS binding near key lysine residues, supporting epitope masking as a key mechanism of reduced IgE recognition. This study provides molecular-level insight into CP-assisted glycation as a targeted strategy for mitigating milk protein allergenicity.