title: Quantitation of human milk proteins and their glycoforms using multiple reaction monitoring (MRM).
abstract: Human milk plays a substantial role in the child growth, development and determines their nutritional and health status. Despite the importance of the proteins and glycoproteins in human milk, very little quantitative information especially on their site-specific glycosylation is known. As more functions of milk proteins and other components continue to emerge, their fine-detailed quantitative information is becoming a key factor in milk research efforts. The present work utilizes a sensitive label-free MRM method to quantify seven milk proteins (α-lactalbumin, lactoferrin, secretory immunoglobulin A, immunoglobulin G, immunoglobulin M, α1-antitrypsin, and lysozyme) using their unique peptides while at the same time, quantifying their site-specific N-glycosylation relative to the protein abundance. The method is highly reproducible, has low limit of quantitation, and accounts for differences in glycosylation due to variations in protein amounts. The method described here expands our knowledge about human milk proteins and provides vital details that could be used in monitoring the health of the infant and even the mother. Graphical Abstract The glycopeptides EICs generated from QQQ.
authors: Department of Chemistry, University of California, Davis, CA, 95616, USA.|Foods for Health Institute, University of California, 1 Shields Ave, Davis, CA, 95616, USA.
authors: Jincui
authors: J
authors: Huang
authors: Department of Chemistry, University of California, Davis, CA, 95616, USA.
authors: Muchena J
authors: MJ
authors: Kailemia
authors: Department of Chemistry, University of California, Davis, CA, 95616, USA.
authors: Elisha
authors: E
authors: Goonatilleke
authors: Department of Chemistry, University of California, Davis, CA, 95616, USA.
authors: Evan A
authors: EA
authors: Parker
authors: Department of Chemistry, University of California, Davis, CA, 95616, USA.|Foods for Health Institute, University of California, 1 Shields Ave, Davis, CA, 95616, USA.
authors: Qiuting
authors: Q
authors: Hong
authors: Department of Drug Chemistry and Technologies, University of Rome, Piazzale Aldo Moro 5, 00185, Rome, Italy.
authors: Rocchina
authors: R
authors: Sabia
authors: Foods for Health Institute, University of California, 1 Shields Ave, Davis, CA, 95616, USA.|Department of Food Science and Technology, University of California, 392 Old Davis Road, Davis, CA, 95616, USA.
authors: Jennifer T
authors: JT
authors: Smilowitz
authors: Foods for Health Institute, University of California, 1 Shields Ave, Davis, CA, 95616, USA.|Department of Food Science and Technology, University of California, 392 Old Davis Road, Davis, CA, 95616, USA.
authors: J Bruce
authors: JB
authors: German
authors: Department of Chemistry, University of California, Davis, CA, 95616, USA. cblebrilla@ucdavis.edu.|Foods for Health Institute, University of California, 1 Shields Ave, Davis, CA, 95616, USA. cblebrilla@ucdavis.edu.
authors: Carlito B
authors: CB
authors: Lebrilla
mesh_terms: D005504:Food Analysis / Q000379:methods*; D006031:Glycosylation; D006801:Humans; D013058:Mass Spectrometry; D008894:Milk Proteins / Q000032:analysis* / Q000737:chemistry*; D008895:Milk, Human / Q000737:chemistry*
chemical_list: D008894:Milk Proteins
keywords: Glycoproteomics; Human milk; MRM; Mass spectrometry; UPLC