CD BioGlyco provides various technologies for HMO Profiling. All these methods provide detailed insights into the oligosaccharide pattern of individual milk samples, frequently paired with further information about the relative amount of single isomers. We have developed systematic solutions to provide clients with One-stop Solution for HMO Production.
Prebiotics, anti-adhesion agents, antibacterial agents, and immunomodulators are biological functions of human milk oligosaccharides (HMO). Characterizing the composition of the specific HMO isolated is essential to investigate the molecular mechanisms involved. Understanding the compositional differences of HMO requires compositional analysis of the entire HMO mixture. However, the diversity and complexity of the structures pose a challenge to their isolation and detection.
A variety of techniques have been demonstrated for the analysis of HMO. These technologies mainly include nuclear magnetic resonance (NMR), mass spectrometer (MS), and liquid chromatography (LC). Among them, NMR and MS are the main methods for studying the HMO structure. LC and capillary electrophoresis combined with UV adsorption or laser-induced fluorescence detection are the main methods for quantifying HMO by labeling natural oligosaccharides with chromophores and fluorophores. However, these methods cannot be used to obtain the structural information of HMO. Therefore, the combination of separation and MS has been widely used to analyze the oligosaccharides of samples.
Fig.1 HMO concentrations (A, C) and relative composition (B) among 427 Canadian mothers in the Canadian Healthy Infant Longitudinal Development cohort, overall (A, B) and by Secretor status (C). (Azad, et al., 2018)
For the identification of HMO, various methods such as paper chromatography, HPLC coupled with MS, high-performance anion exchange chromatography (HPAEC-PAD), HPLC chip MS, capillary gel electrophoresis/laser-induced fluorescence techniques, or LC-HPLC-MS have recently been used.
CD BioGlyco provides a variety of effective methods for compositional analysis. Taking LC-MS analysis as an example, our analytical strategy is as follows.
A crude mixture of HMOs is prepared from pooled human milk samples. The refined HMO mixture is separated from residual peptides and a large amount of lactose.
Derivatization of HMOs is divided into three steps. (a) Labeling by reductive amination. (b) Labeling by Michael addition. (c) Derivatization by reduction.
Conditions of ionic strength, pH, and buffer system are optimized to achieve robust elution behavior for LC and sensitive ionization efficiency for MS.
Analysis of the resulting data uses statistical principles.
Fig.2 Schematic of the sample preparation process for a comprehensive comparison. (Lang, et al., 2021)
CD BioGlyco is committed to providing the best solutions for global clients in the field of glycoscience. We guarantee to provide satisfactory results of HMO compositional profiling for clients all over the world. If you are interested in our services, please contact us for more details without any hesitation.
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