A5016547487- Polysaccharides and Plant Cell Walls
- Microbial Metabolites in Food Biotechnology
- Glycosylation and Glycoproteins Research
- Infant Nutrition and Health
- Advanced Proteomics Techniques and Applications
- Polysaccharides Composition and Applications
- Enzyme Production and Characterization
- Mass Spectrometry Techniques and Applications
- Plant nutrient uptake and metabolism
- Biosensors and Analytical Detection
- Carbohydrate Chemistry and Synthesis
- Biofuel production and bioconversion
- Monoclonal and Polyclonal Antibodies Research
- Postharvest Quality and Shelf Life Management
- Digestive system and related health
- SARS-CoV-2 and COVID-19 Research
- Analytical Chemistry and Chromatography
- Fungal Biology and Applications
- Influenza Virus Research Studies
- Animal Nutrition and Physiology
- Biochemical Analysis and Sensing Techniques
- Electrospun Nanofibers in Biomedical Applications
- Food composition and properties
- SARS-CoV-2 detection and testing
- Probiotics and Fermented Foods
National Institute of Standards and Technology
2018-2024
Material Measurement Laboratory
2018-2023
Wageningen University & Research
2012-2019
National Institute of Standards
2019
Pectic oligosaccharides (POS) were obtained by hydrothermal treatment of orange peel wastes (OPW) and purified membrane filtration to yield a refined product containing 90 wt % the target products. AraOS (DP 3-21), GalOS 5-12), OGalA 2-12, with variable DM) identified in POS mixtures, but long-chain products also present. The prebiotic potential concentrate was assessed vitro fermentation using human fecal inocula. For comparative purposes, similar experiments performed pectin commercial...
We report the development and availability of a mass spectral reference library for oligosaccharides in human milk. This represents new variety that includes consensus spectra compounds annotated through various data analysis methods, concept can be extended to other varieties biological fluids. Oligosaccharides from NIST Standard Reference Material (SRM) 1953, composed milk pooled 100 breastfeeding mothers, were identified characterized using hydrophilic interaction liquid chromatography...
This study significantly expands both the scope and method of identification for construction a previously reported tandem mass spectral library 74 human milk oligosaccharides (HMOs) derived from results combined LC-MS/MS experiments comprehensive structural analysis HMOs. In present work, hybrid search "bootstrap" was employed that substantially broadens coverage thereby increases utility use spectrum library-based rapid tentative all distinguishable glycans in milk. involved searching...
We report the comparison of mass-spectral-based abundances tryptic glycopeptides to fluorescence released labeled glycans and effects mass charge state in-source fragmentation on glycopeptide abundances. The primary glycoforms derived from Rituximab, NISTmAb, Evolocumab, Infliximab were high-mannose biantennary complex galactosylated fucosylated N-glycans. Except for ions loss HexNAc or HexNAc-Hex sugars are prominent other therapeutic IgGs. After excluding results, a linear correlation was...
Glycopeptide Abundance Distribution Spectra (GADS) were recently introduced as a means of representing, storing, and comparing glycan profiles intact glycopeptides. Here, using that representation, an extensive analysis is made multiple commercial sources the recombinant SARS-CoV-2 spike protein, each containing 22 N-linked sites (sequons). Multiple proteases are used along with variable energy fragmentation followed by ion trap confirmation. This enables detailed examination reproducibility...
A method for representing and comparing distributions of N-linked glycans located at specific sites on proteins is presented. The representation takes the form a simple mass spectrum given peptide sequence, with each peak corresponding to different glycopeptide. (in place m/z) that glycan mass, its abundance corresponds relative in electrospray MS1 spectrum. This provides facile means all identifiable glycopeptides arising from single protein "sequon" thereby enabling comparison searching...
Annotating product ion peaks in tandem mass spectra is essential for evaluating spectral quality and validating peptide identification. This task more complex glycopeptides crucial the confident determination of glycosylation sites glycoproteins. MS_Piano (Mass Spectrum Peptide Annotation) software was developed reliable annotation collision induced dissociation (CID) peptides or N-glycopeptides given sequences, charge states, optional modifications. The program annotates each peak high low...
We present a mass spectral library-based method for analyzing site-specific N-linked protein glycosylation. Its operation and utility are illustrated by applying it to both newly measured available proteomics data of human milk glycoproteins. It generates two varieties libraries. One contains glycopeptide abundance distribution spectra (GADS). The other tandem the underlying glycopeptides. Both originate from identified glycopeptides in proteolytic digests purified glycoproteins, which...
This work presents methods for identifying and then creating a mass spectral library disulfide-linked peptides originating from the NISTmAb, reference material of humanized IgG1k monoclonal antibody (RM 8671). Analyses involved both partially reduced non-reduced samples under neutral weakly basic conditions followed by nanoflow liquid chromatography tandem spectrometry (LC-MS/MS). Spectra containing disulfide bonds are identified MS1 ion MS2 fragment data in order to completely map all...
This work presents a detailed determination of site-specific N-glycan distributions the recombinant influenza glycoproteins hemagglutinin and neuraminidase. Variation in glycosylation among is not predictable can depend on details biomanufacturing process as well protein structure. In this study, proteins were analyzed from eight strains four different suppliers. These include five three neuraminidase proteins, each produced HEK293 cell line. Digestion was conducted using series complex...
Mammalian milk oligosaccharides (MMOs) from human and farm animals have been of great interest because their functional application in food nutrition. However, the biological significance variation proportions MMOs among milks is poorly understood. MMO concentration (0.05 - 0.3 g/L) mature animal >20 fold lower than (HMOs) (12 [1]. Hence, screening techniques are necessary for structural characterization milk. This study characterizes compares chemical structures derived tandem mass spectra...