Frequently Asked Questions

What Glyko products do I need for glycan profiling of an antibody by HPLC using 2-AB fluorescent label?

We recommend:

• GKE-5010 N-Glycanase® Plus for deglycosylation of the Mab. See FAQ on our website at:
  http://www.prozyme.com/faqs/gke-5010faq.pdf Product listing and tech sheet: http://www.prozyme.com/pdfs/gke-5010.pdf
  
• GKI-4756 GlycoClean™ R Cartridge, optional for cleanup of the glycan samples (protein removal) if you have problems with the ethanol precipitation, due to the limited amounts of the protein or its solubility in ethanol:
  http://www.prozyme.com/glyko/columns.html#glycoclean
The product insert is posted on our website:
http://www.prozyme.com/pdfs/gki-4756.pdf
• GKK-404 Signal™ 2-AB Labeling Kit for labeling the glycans:
  http://www.prozyme.com/glyko/signal.html#gkk-404
The product insert is posted on our website:
http://www.prozyme.com/pdfs/gkk-404.pdf
• GKI-4726 GlycoClean S Cartridges are used after 2-AB labeling to remove unreacted dye and salts:
  http://www.prozyme.com/glyko/columns.html#glycoclean
  The product insert is posted on our website:
  http://www.prozyme.com/pdfs/gki-4726.pdf
• GKI-4728 GlycoSep™ N Column with the GKI-4728G N Guard Column are used to separate the 2-AB-labeled glycans from one another (mostly by size). The product insert is posted to our website:
  http://www.prozyme.com/pdfs/gki-4728.pdf
• GKSB-503 2-AB-(Glucose Homopolymer) Standard is used to confirm proper functioning of the HPLC column, and to calculate GU numbers to corroborate and identify peaks.
  http://www.prozyme.com/pdfs/c-a/gksb-503-2-ab.pdf
• 2-AB-labeled Standards are available for glycan identification:
  http://www.prozyme.com/glyko/carbostandards.html#2-ab
• Various glycan standards appropriate for monoclonals are:
  
  • NGA2, NGA2F, NA2, NA2F, A2 and A2F. If the MAb is from mouse hybridoma, add NA2FB.
  • NGA2 (aka G0) - GKSB-301 2-AB-Asialo-, agalacto-, biantennary
  • NGA2F - GKSB-302 2-AB-Asialo-, agalacto-, biantennary, core-substituted with fucose
  • NA2 (aka G2) - GKSB-304 2-AB-Asialo-, galactosylated biantennary
  • NA2F - GKSB-305 2-AB-Asialo-, galactosylated biantennary, core-substituted with fucose
  • A2 - GKSB-312 2-AB-Di-sialylated-, galactosylated biantennary
  • A2F - GKSB-313 2-AB-Di-sialylated-, galactosylated biantennary, core-substituted with fucose
  • NA2FB - GKSB-306 2-AB-Asialo-, galactosylated biantennary, core-substituted with fucose and with bisecting GlcNAc
• These standards are also available unlabeled for validation of the 2-AB labeling step. These are also useful as standards for analysis of the cleaved glycans with MALDI-TOF mass spectrometry:
  http://www.prozyme.com/glyko/carbostandards.html#n-linked-oligo
  
  • NGA2 (aka G0) - GKC-004300 Asialo-, agalacto-, biantennary
  • NGA2F - GKC-004301 Asialo-, agalacto-, biantennary, core-substituted with fucose
  • NA2 (aka G2) - GKC-024300 Asialo-, galactosylated biantennary
  • NA2F - GKC-024301 Asialo-, galactosylated biantennary, core-substituted with fucose
  • A2 - GKC-224300 Di-sialylated-, galactosylated biantennary
  • A2F - GKC-224301 Di-sialylated-, galactosylated biantennary, core-substituted with fucose
  • NA2FB - GKC-024311 Asialo-, galactosylated biantennary, core-substituted with fucose and with bisecting GlcNAc
• GKI-4025 GlycoClean H Cartridges for glycan cleanup when MALDI-TOF is employed for analysis. H cartridges remove buffer salts in the samples.
  http://www.prozyme.com/pdfs/gki-4025.pdf

N-Glycanase® is a registered trademark of Genzyme Corporation, Boston, MA, USA. Recombinant PNGase F is covered by US Patent No 5,238,821, which is exclusively licensed for manufacture by ProZyme, Inc.