Glycobiology

The mAbsolute kits, are low- and high-throughput glycan analysis kits, based on highly pure synthetic isotope-labeled internal standards for the absolute quantification of mAb glycosylation by MALDI-TOF MS. Minimal handling and time are required for sample preparation and within seconds, all relevant glycans of a deglycosylated mAb sample are quantified by comparison of MALDI profiles with the corresponding internal reference standards.

CarboQuant Glycan Standards are patented synthetic 13C-labeled N-glycans, produced entirely in our laboratory for use as internal standards in absolute glycan quantification by mass spectrometry. They are available as single standards or in kit form including a quantification software and all reagents and consumables required for glycan analysis.

Other applications in the field of glycobiology research and proteomics:

  • Increase reproducibility in lab-to-lab method transfer (internal calibration standard)
  • De-convolute and quantify co-eluting peaks in LC-MS
  • Quantify glycan recovery after sample preparation

CarboClip is a recombinant peptide-N-Glycosidase F (PNGase F), from Elizabethkingia sp, overexpressed in E. coli that contains a hexahistidine tag in its C-term and cleaves between the GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides from N-linked glycoproteins. CarboClip is provided glycerol-free as a lyophilized powder containing Tris-HCl, NaCl2, and Na2EDTA.

Activity: After reconstitution in 150 μL of UltraPure H2O, 1 μl of CarboClip can deglycosylate > 95% of ribonuclease B (1 mg) in less than 20 minutes. Activity is calculated on a 12 % SDS-PAGE and percent of deglycosylation using densitometry.

Storage: Lyophilized powder: -80 °C and -20 °C >1 year. At room temperature 10 days. Reconstituted protein should be stored at 4 °C and is stable for 2-3 months

Characterization of glycosylation in proteins, antibodies, hormones, and other glycoconjugates using lectin microarray

Glycans are detached from the glycosylated protein using PNGase F (CarboClip) and occupancy is determined by comparing the abundance of deamidated asparagine against the one containing aspartic acid (non glycosylated)

Quick determination of the glycosylation profile of a expressed recombinant protein depending on the heterologous host. Correlation of the differences in glycosylation and protein function

MALDI-TOF MS based glycan structure identification, including α-2,3/ α-2,6 differential sialylation following a sialic acid stabilization procedure and using CarboQuant glycan standards

N-acetyl and N-glycolyl neuraminic acid (NANA and NGNA) quantification by RPLC, using DMB labeling.

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Proteomics

Qualitative and quantitative determination of the amino acid composition of antibodies, proteins, peptides and others for research and pharmaceutical applications

  • Clinical research in Universities and veterinary labs.
  • Quality Control and compositional analysis in biosciences and pharmaceutical industries.

Measurement of absolute heat capacities of biopolymers in dilute solution

  • Biopolymer Solution Conformation & Solvation (Absolute heat capacities)
  • Biopolymer Stability (Protein denaturation)
  • Biopolymer Structure (Domain organization)
  • Bioengineering (Mutant proteins)
  • Ligand Interactions (Drug binding to proteins or nucleic acids)
  • Membrane Structure (Lipid Bilayers, membrane proteins)
  • Polynucleotides (Helix to coil transitions).

Deglycosylation systematically improves N-glycoprotein identification in liquid chromatography–tandem mass spectrometry proteomics

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