mAbsolute kits have been optimized for the MALDI-TOF based absolute glycan quantification using Bruker Autoflex and UltrafleXtreme III equipments. Validation is on-going for the application of the kits to different MALDI-TOF instruments and to other mass spectrometry techniques.
Video shows how to proceed to analyze glycan content of mAb using mAbsolute Kit.
each kit contains
- Quantified mixture of 8-11 13C-standards covering all major glycans present in therapeutic monoclonal antibodies expressed in CHO cell lines. Complete kit also includes 2,3-sialylated standards. Also, a humanized mixture of standards has been developed covering also bisected-biantennary standards as well as 2,6-sialylated structures, for applications in human IgG glysosylation analysis, both Fc and Fab regions.
- CarboClip enzyme: Proprietary peptide N-glycosidase F (PNGase F). The glycan release from the proteic backbone is performed by CarboClip, an enzyme that cleaves the amide bond between the N-glycan and the asparagine residue in the protein. The enzyme included in the kit has been developed in-house and is provided as lyophilized powder to maintain a high deglycosylation activity.
- Sialic acid stabilization procedure. Despite the mild ionization conditions of MALDI-TOF techniques, the sialylated N-glycans can be partially hydrolized during the laser induced-desorption step concerning the quantification of the sample. A simple derivatization step included in the sample preparation protocol avoids the desialylation and also helps distinguising the different 2,3- and 2,6-sialyl regioisomers, that can be present in a sample.
- Proprietary Quantification Software. The MALDI-TOF based quantification method is compatible with a high-troughput performance, yielding a complex data set which can be treated easily with the quantification software provided with the kit. Despite it was developed for Bruker data formats, it is very simple to import data in general and common formats as .csv, analyzing tenths or hundreds of experiments automatically. The software generates also a customized report regarding absolute and relative quantification data.
User Manuals for the mAbsolute Kits:
mAbsolute Neutral Kit _ User Manual
mAbsolute Complete Kit _ User Manual
mAbsolute Human serum IgG Kit _ User Manual
mAbsolute Kits find application on:
- Rapid Absolute glycan quantification
- Measure of glycan recovery for in sample preparation
- Internal standards for the quantitative glycan profiling by MALDI-Tof MS employing mAbsolute kits
- Rapid high-throughput glycan analysis in clone selection and process development
- Quality control
- Biosimilar optimisation
Clinical Chemistry Biomarkers
- Measure glycan levels in serum samples with picomole precision
- Establish cut-off values for glycan biomarkers
- Quantify aberrant glycosylation in cancer samples
- Internal performance calibrants for method transfer and multiple laboratory studies