Fast Glycan Labeling & Analysis: Sensitive CE-LIF Detection with Automated Glycan Identification
This webinar discusses a Fast Glycan Labeling & Analysis technology which has been developed for N-linked oligosaccharide profiling of glycoproteins along with a gel-buffer system to ensure rapid and high-resolution separation of the target molecules. Glycan release, fluorophore labeling and clean-up parameters were all optimized resulting in 60 min sample preparation time using a novel magnetic bead mediated protocol that assures excellent yield, high reproducibility and easy automation. Optimization included rapid endoglycosidase digestion and fluorophore labeling time and temperature as well as sample clean-up, supporting fast sample processing. The procedure does not require any centrifugation and vacuum centrifugation steps, otherwise necessary for most glycan sample preparation methods.
Capillary electrophoresis analysis with laser induced fluorescence detection (CE-LIF) of the fluorophore (APTS) labeled glycans was also optimized to enable rapid and high resolution separations of the labeled sugars just within a few minutes, in order to accommodate both manual and automated sample processing. We also report on the design and implementation of a co-injection based triple-internal standard mediated glycoinformatics method, to alleviate the need of an accompanying run of the maltooligosaccharide ladder for precise glucose unit (GU) calculation based structural assignment. The importance of precise temperature control during CE-LIF analysis is also emphasized. Worked examples will show comprehensive glycan composition analysis of some of the most prescribed monoclonal antibody therapeutics and their biosimilar counterparts. Quantitative assessment and automatic structural identification of glycans of CQA importance in biologics development will also be discussed.
In this webinar, you will learn:
How to run glycan samples up to 5x faster than HILIC
60 minute sample prep for up to 96 glycan samples
How to separate and identify N-glycans in 5 minutes