SmartEnzymes Poster Highlights 2019

Which are the top five poster from Genovis in 2019? We have selected the posters that are both popular with our website visitors and that describe the most exciting new applications using SmartEnzymes. Click on the images to download the full posters.

1) ASMS Poster, 2019

1 – Analysis of O-glycosylated-Biopharmaceuticals using an O-glycan Dependent Endoprotease and LC-MS (ASMS, 2019)

In this collaborative work, we set out to combine our novel, specific enzymes with the latest LC-MS technologies from Thermo Fisher Scientific in order to improve and simplify analytical workflows for biotherapeutics. We demonstrate in-depth characterization of O-glycosylated biopharmaceuticals and quantitative comparison of O-glycosylation patterns. We also present a workflow for total deglycosylation of heavily glycosylated biopharmaceuticals, allowing for intact mass spectrometry analysis without interference from glycan heterogeneity.

2) World ADC Poster, 2019

2 – Robust Generation of Site-specific and Homogeneous Antibody Conjugates using GlyCLICK® (World ADC, 2019)

In this work, we present details on the enzymatic processing of the Fc-glycans that result in the homogenous conjugates, and confirmed it by mass spectrometry. The immunoreactivity of the conjugated antibodies was studied using surface plasmon resonance and toxicity by a dose-dependent response of a DM1 GlyCLICK conjugated trastuzumab (T-DM1).

CASSS AT Poster, 2019

3 – An Automated Workflow for Analysis of Monoclonal Antibody Subunits (CASSS AT 2019)

Here we present a rapid, automated solution for antibody subunit generation and analysis using a standard HPLC-MS setup with only minor modifications. FabRICATOR® (IdeS) enzyme was immobilized in an HPLC column format to allow for easy on-column digestion of IgG-based biologics followed by RP-HPLC-MS analysis. This facilitates a fully automated, completely hands-off workflow for analysis of several critical quality attributes.

4 – FabALACTICA® Generates Pure and Homogenous mAb Subunits that Facilitate 2D-NMR Spectroscopy Analysis (Festival of Biologics, 2019)

4) Festival of Biologics Poster, 2019

Two-dimensional nuclear magnetic resonance spectroscopy (2D-NMR) allows for the precise atomic-level comparison of higher order structure (HOS) for IgG-based biopharmaceuticals. Since most of the approved therapeutic mAbs today have a human IgG1 backbone, the cysteine protease FabALACTICA (IgdE) can simplify the analysis. The enzyme digests human IgG1 at a specific site above the hinge, generating intact Fab and Fc fragments. In this work, we present a workflow for obtaining homogeneous Fab and Fc fragments that are ideal for evaluating HOS of the chimeric mAb infliximab using 2D-NMR.

5 – Complete and Rapid Desialylation of Therapeutic Glycoproteins using Immobilized SialEXO® (AET 2019)

The enzymatic performance of the Immobilized SialEXO column was tested on therapeutic glycoprotein substrates: human C1 inhibitor, etanercept, cetuximab, and human tissue-type plasminogen activator (tPA). Treated and native glycoproteins were then analyzed using released glycan analysis, antibody subunit LC-MS, and capillary iso-electric focusing. The sialidase column delivered complete de-sialylation of 0.5 mg glycoprotein after 30 min at room temperature. In summary, the new Immobilized SialEXO column provides robust and rapid desialylation ideal for routine analysis of biopharmaceuticals with a range of commonly used analytical techniques

AET Poster, 2019