title: Development of a high-throughput glycoanalysis method for the characterization of oligosaccharides in human milk utilizing multiplexed capillary gel electrophoresis with laser-induced fluorescence detection.
abstract: During the last decade, enormous progress regarding knowledge about composition and properties of human milk (HM) has been made. Besides nutrition, the three macro-nutrients: proteins, lipids, and carbohydrates combine a large variety of properties and functions. Especially, complex oligosaccharides emerge as important dietary factors during early life with multiple functions. The characterization of these HM oligosaccharides (HMOS) within the total carbohydrate fraction is prerequisite to understand the relationship between milk composition and biological effects. Therefore, extended studies of large donor cohorts and thus, new high-throughput glycoanalytical methods are needed. The developed method comprises sample preparation, as well as analysis of HMOS by multiplexed CGE with LIF detection (xCGE-LIF). Via a respective database the generated "fingerprints" (normalized electropherograms) could be used for structural elucidation of HMOS. The method was tested on HM samples from five different donors, partly sampled as a series of lactation time points. HMOS could be easily identified and quantified. Consequently, secretor and Lewis status of the donors could be determined, milk typing could be performed and quantitative changes could be monitored along lactation time course. The developed xCGE-LIF based "real" high-throughput HMOS analysis method enables qualitative and quantitative high-performance profiling of the total carbohydrate fraction composition of large sets of samples.
authors.affiliation: Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Kottler
authors.forename: Marko
authors.initials: M
authors.lastname: Mank
authors.forename: René
authors.initials: R
authors.lastname: Hennig
authors.forename: Beate
authors.initials: B
authors.lastname: Müller-Werner
authors.forename: Bernd
authors.initials: B
authors.lastname: Stahl
authors.forename: Udo
authors.initials: U
authors.lastname: Reichl
authors.forename: Erdmann
authors.initials: E
authors.lastname: Rapp
mesh_terms: D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D062126:Databases, Chemical; D019075:Electrophoresis, Capillary / Q000379:methods*; D005260:Female; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008895:Milk, Human / Q000737:chemistry*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry / Q000145:classification; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D009844:Oligosaccharides
keywords: CGE-LIF; Fingerprint; Human milk; Oligosaccharides