New references on IgG glycosylation, glycation and ADC characterization using IgGZERO and FabRICATOR

March 23, 2016 | References |

New references are out using Genovis enzymes to study antibody glycation, pairing of high-mannose glycans and ADC characterization using CE-MS.


Effect of high mannose glycan pairing on IgG antibody clearance.

In a paper published in the journal Biologics, researchers from Amgen in California studied the pairing of Fc glycans on a therapeutic monoclonal antibody. This is an area of immense interest, as most glycan analysis approaches assume that all glycoforms are distributed equally. Liu et al. used the limited activity of IgGZERO on high-mannose glycans to study symmetrical or assymetrical high-mannose on a mAb from a pharmacokinetics study. The results showed that high-mannose glycans are cleared from circulation independent on the high-mannose symmetry.


Glycation of polyclonal IgGs: Effect of sugar excipients during stability studies

Researchers at LFB Biotechnologies have studied glycation of the Fc/2 fragment during stability studies using LC/MS instrumentation. For 6 months an IVIG preparation was kept at various temperatures and for analysis an LC/MS method using digestion of the IVIG with FabRICATOR and deglycosylation using IgGZERO, the levels of glycation was studied. It was found that high levels of glycation from reducing sugars such as glucose and maltose were detected in the first month of stability at 25°C and that their reactivity was lower at elevated temperatures.


Structural characterization of antibody drug conjugate by a combination of intact, middle-up and bottom-up techniques using sheathless capillary electrophoresis – tandem mass spectrometry as nanoESI infusion platform and separation method.

In a report using sheatless CE-MS to characterize a ADC molecule (Brentuximab vedotin), FabRICATOR was used to digest the antibody and generate specific subunits. The ADC was studied at multiple levels, including intact, middle-down (FabRICATOR) and bottom-up. The limited digestion using FabRICATOR allowed characterization of the drug load of the individual subunits. The authors claim this approach reduced the number of experiments required for full characterization of the ADC.