Identification of Glycosylation-Associated Proteins in Colostrum: A Review of Mass Spectrometry-Based Strategies.

Colostrum, first milk produced by mammalians after parturition, is a biologically rich and complex fluid, referred to as "liquid gold" due to its exceptional nutritional and immunological properties. Among the bioactive molecules, proteins play critical roles in neonatal development and immune protection, notably due to the presence of glycosylations which support protein folding, stability, recognition, and host-microbe interactions. Thus, glyco-associated proteome characterization in colostrum by mass spectrometry is plebiscite to elucidate its biological functions and therapeutic potential. In this review, we provide a comprehensive overview of MS-based workflows employed for colostrum glycoproteins analysis, highlighting commonly used strategies, methodological specificities, and the gathered biological information. We also discuss alternative analytical approaches that have been successfully applied to colostrum glycoprotein studies. Finally, we present a critical perspective on emerging technologies and unexplored workflows, evaluating their potential applicability, limitations, and future relevance for the analysis of glycosylation-associated proteins in colostrum.