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HALO Glycan
The Halo Glycan Columns from Advanced Materials Technology are the newest member of the Halo Fused BioClass family. They are especially designed for bioseparations of oligosaccharides and particularly protein-linked glycans and glycopeptides by HILIC, H(ydrophilic) I(interactive) LI(quid) C(hromatography).
Chemistry:
- Highly polar ligand with 5 hydroxyl groups
- Ligand tethered to 2.7µm Fused Core silica via novel proprietary linkage chemistry
- Fused-Core particle with 135 m2/g surface area and 90A pore size
For separations of:
- Oligosaccharides
- Protein-linked glycans andglycopeptides
- Example application: RNGase-Released and Labeled N-Glycans (see right)
For Quality Assurance QA every lot is tested by separation of a procainamide-reducing end-labeled glycan ladder of oligosaccharides of 2-25 glucose units. The peaks of 5 and 10 glucose units have to meet strict specifications of retention and peak widths before the lot is approved for the HALO Glycan columns.
Applications
Separation of Procainamide-Labeled Dextran Standards on HALO Glycan
A HALO Glycan column shows an efficient separation of procainamide-labeled dextran standards (Sigma-Aldrich 1:1 (w/w) of Part numbers 00268 and 00269) at 0.5 µg/µL in 70% ACN/30% water. Each lot of HALO Glycan packing is tested using this sample to assure lotto-lot reproducibility and performance.
TEST CONDITIONS
- Column: 2.1 x 150 mm, HALO 2.7 μm Glycan
- Part Number: 92922-705
- Mobile Phase A: 50 mM Ammonium Formate, pH 4.45
- Mobile Phase B: Acetonitrile
- Gradient: 80-55% B in 25 min
- Flow Rate: 0.6 mL/min.
- Pressure: 190 bar
- Temperature: 60°C
- Detection: UV 300 nm
- Injection Volume: 3 μL Sample
- Solvent: 70/30 ACN/water
- Response Time: 0.5 sec.
- Data Rate: 3.3 Hz
- Flow Cell: 2.5 μL semi-micro
- LC System: Shimadzu Nexera
Separation of PNGase-Released and Labeled N-Glycans By HILIC Using HALO Glycan Column
A fast separation of PNGase-released and procainamidelabeled N-Glycans from Ribonuclease B is accomplished with a HALO Glycan column.
PEAK IDENTITIES:
- 1. PAm-GlcNAc2Man5
- 2. PAm-GlcNAc2Man6
- 3. PAm-GlcNAc2Man7
- 4. PAm-GlcNAc2Man8
- 5. PAm-GlcNAc2Man9
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