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.
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