Course 2
SEPARATION AND CHARACTERIZATION OF GLYCOPROTEIN AND GLYCOLIPID OLIGOSACCHARIDES (August 10-14, 2026)
Monday, August 10, 2026
8:45 a.m. – 9:00 a.m.
Introduction and Welcome
Dr. Parastoo Azadi
9:00 a.m. – 10:00 a.m.
Lecture – “Overview of Glycoprotein Structures, Biosynthesis and Function”
Dr. Kelley Moremen
10:00 a.m. – 10:15 a.m. – Break
10:15 a.m. – 11:15 a.m.
Lecture continued – “Overview of Glycoprotein Structures, Biosynthesis and Function”
Dr. Kelley Moremen
11:15 a.m. – 12:00 p.m.
Lab Exercise discussion
SECTION I – Monosaccharide composition analysis by HPAEC-PAD
Laboratory
SECTION I – Monosaccharide composition analysis
Begin acid hydrolysis
Dr. Varughese (Alex) Mulamoottil
12:00 p.m. – 1:00 p.m. – Lunch
1:00 p.m. – 2:00 p.m.
Lecture –“Labeling and Separation of Carbohydrates”
Dr. Stephanie Archer-Hartmann
2.00 p.m. – 2.45 p.m.
Lab Exercise discussion
SECTION IV – Permethylation of glycans
Drs. Sonon/Kumar
2:45 p.m. – 3:00 p.m. -Break
3:00 p.m. – 3:30 p.m.
Laboratory
SECTION I – Monosaccharide composition analysis
Recover samples from hydrolysis, freeze digests and lyophilize
Dr. Varughese (Alex) Mulamoottil
SECTION II – Release of N-linked glycans from a glycoprotein (Fetuin)
Denature glycoprotein and start trypsin digestion
Drs. Sonon/ Kumar
3:30 p.m. – 4:15 p.m.
Lab exercise discussion
SECTION VI – Separation of glycolipids by TLC
Laboratory
SECTION VI – TLC analysis
Desialylation of porcine brain gangliosides
Dr. Stephanie Archer-Hartmann
4:15 p.m. – 5:00 p.m.
Lab Exercise discussion
SECTION III – Release of O-glycans from a glycoprotein (Mucin)
Laboratory
SECTION III – Release of O-glycans
β-elimination of O-linked glycans
Drs. Sonon/ Kumar
Tuesday, August 11, 2026
9:00 a.m. – 10:00 a.m.
Lecture – “Glycan and Glycoprotein Analysis”
Dr. Parastoo Azadi
10:00 a.m. – 12:00 p.m.
Laboratory
SECTION II – Release of N-glycans
Terminate Trypsin digestion of Fetuin
Drs. Sonon/Bhoj Kumar
Release of N-glycans
Release N-glycans with PNGase F
Drs. Sonon/ Kumar
SECTION III – Release of O-glycans
Neutralization and de-salting of beta-eliminated O-linked glycans
Drs. Sonon/Bhoj Kumar
SECTION VI – TLC analysis
Sample cleaning by C18 reversed phase cartridge
Dry sample under N2
Dr. Stephanie Archer-Hartmann
12:00 p.m. – 1:00 p.m. – LUNCH
1:00 p.m. – 2:15 p.m.
Lecture – “Bacterial Glycoproteins”
Dr. Christine Szymanski
2:15 p.m. – 3:00 p.m. – CCRC Tour/Break
Dr. Christian Heiss
3:00 p.m. – 3:30 p.m. –
Laboratory
SECTION I – Monosaccharide composition analysis
Preparation of monosaccharide digests for HPAEC analysis
Dr. Varughese (Alex) Mulamoottil
3:30 p.m. – 4:45 p.m.
Lecture – “Applications of Ion Mobility in the Analysis of N-Linked Glycans”
Dr. Ron Orlando
4:45 p.m. – 5:00 p.m.
Lab exercise discussion
SECTION V – Labeling of free N-glycans by Reductive Amination
Laboratory
SECTION V – Procainamide labeling
Dr. Stephanie Archer-Hartmann
Wednesday, August 12, 2026
9:00 a.m. – 10:00 a.m.
Lecture – “Glycans Linked to Lipids and Lipid Precursors”
Dr. Michael Tiemeyer
10:00 a.m. – 10:10 a.m. – Break
10:10 a.m. – 11:10 a.m.
Lecture – “Introduction to HPAEC”
Dr. Parastoo Azadi
11:10 a.m. – 12:00 p.m.
Laboratory
SECTION II – Release of N-glycans
Separation of N-glycans from O-glycopeptides/peptides by C18 sep pak
Drs. Sonon/ Kumar
12:00 p.m. – 1:00 p.m. – LUNCH
1:00 p.m. – 2:00 p.m.
Laboratory
SECTION III – Release of O-glycans
Removal of Borates from beta-eliminated O-linked glycans
Dr. Bhoj Kumar
2:00 p.m. – 2:30 p.m.
SECTION I – HPAEC Data Discussion
Dr. Varughese (Alex) Mulamoottil
2:30 p.m. – 2:40 p.m. – Break
2:40 p.m. – 4:00 p.m.
Laboratory
SECTION VI – TLC analysis
Spot samples
Develop plate
Detection of samples
Dr. Stephanie Archer-Hartmann
4:00 p.m. – 5:00 p.m.
Laboratory
SECTION V – Procainamide Labeling
Removal of free label with SPE cartridges
Lyophilize to dryness.
Dr. Stephanie Archer-Hartmann
Thursday, August 13, 2026
Laboratory
8.45 a.m. – 10:10 a.m.
SECTION IV – Permethylation of released oligosaccharides
(N-glycans from Fetuin or O-glycans from Mucin)
10:10 a.m. – 10:20 a.m. – Break
10:20 a.m. – 11:00 a.m.
Laboratory
SECTION IV – Permethylation (continued)/ Introduction to micropermethylation
Dr. Bhoj Kumar
11:00 a.m. – 12:00 p.m.
Glycan Imaging Mass Spectrometry: theoretical overview and quick demo
Mehrnoush Ghahfarrokhi
12:00 p.m. – 1:00 p.m. – Group Photo and LUNCH
1:00 p.m. – 2:15 p.m.
Laboratory
SECTION V – Procainamide Labeling
Overview of HPLC Chromatography
Data Analysis
Dr. Stephanie Archer-Hartmann
2:15 p.m. – 2:55 p.m.
Demonstration
MALDI TOF/TOF MS demonstration
Dr. Roberto Sonon/Bhoj Kumar
2:55 p.m. – 3:05 p.m. – Break
3:05 p.m. – 3:45 p.m.
HPAEC and HPLC/CE demonstration
Dr. Varughese (Alex) Mulamoottil /Dr. Stephanie Archer-Hartmann
Discussion of data
3:45 p.m. – 4:00 p.m.
SECTION VI – TLC results
Analysis of TLC data
Dr. Stephanie Archer-Hartmann
4:00 p.m. – 5:00 p.m.
SECTION II, III, IV – MALDI TOF/TOF-MS result
Analysis of N-and O-linked glycans MALDI data & Data analysis
Drs. Sonon/ Kumar
Friday, August 14, 2026
<Mass Spectrometry Module>
9:00 a.m. – 12:00 p.m.
Lecture – Glycomics & Glycoproteomics
Dr. Parastoo Azadi
10:15 a.m. – 10:30 a.m. – Break
12:00 p.m. – 1:00 p.m. – LUNCH
1:00 p.m. – 3:00 p.m.:
Mass Spectrometry Facilities Tour – Mass Spec BINGO
MSMS Analysis of Glycans and Glycopeptides
Dr. Stephanie Archer-Hartmann
Or <NMR Module>
9:00 a.m. – 12:00 p.m.
Lecture – “Introduction to NMR of glycoproteins and carbohydrates”
Dr. Christian Heiss
12:00 p.m. – 1:00 p.m. – LUNCH
Afternoon:
Demonstration and data interpretation
Or <Molecular Modeling Module>
9:00 a.m. – 12:00 p.m.
Lecture – “Introduction to Molecular Modeling
Dr. Lachele Foley
12:00 p.m. – 1:00 p.m. – LUNCH
Afternoon:
Demonstration and data interpretation