Science Café Lunch Monday March 27, 12.15-13.15, room: Sorbonne 2, presented by SCIEX
Advances in the Separation of Post-Translational Modifications using CE-MS
Prof. Herbert Lindner, (Innsbruck Medical University, Austria)
In this talk we show how CESI-MS, has been used to detect challenging post-translational modifications including deamidation in crude protein fractions. We will demonstrate how CESI-MS can be used to detect citrullination in intact and digested proteins and in combination with MS/MS and electron transfer dissociation (ETD) fragmentation, locate the presence of deiminated Arginine. Finally, we will discuss how CESI-MS can be used to separate and quantify individual positional isomers of isobaric mono-phosphorylated peptides obtained in the course of a kinase activity study.
Fast Glycan Labeling and Analysis: High-Resolution Separation and Identification in Minutes
Prof. Andras Guttman, (SCIEX, USA)
There is a growing demand in the biopharmaceutical industry for rapid N-glycosylation profiling of biologics. We will present a fast glycan labeling and analysis approach using a novel magnetic bead-mediated process and a CE gel-buffer separation system to quickly profile glycoprotein N-linked carbohydrates. The rapid CE-LIF separation gave excellent yield, high reproducibility, supported large scale 96-well plate sample processing and was easy to automate. A triple-internal standard method significantly speeded up the glycoinformatics tasks and enabled precise glucose unit (GU) assignment of human IgG glycans (relative standard deviation ≤1.07%).
Science Café Lunch Tuesday March 28, 12.15-13.15, room: Sorbonne 2, presented by Agilent Technologies
Solutions for Orthogonal Separations using Electrophoresis and Chromatography and 2D-LC
Dr. Monika Dittmann and Dr. Martin Greiner, (Agilent Technologies, Germany)
Analytical tasks in Biopharma are manifold and complex. In example for characterization of Antibodies trust in a single technique would have a high risk to miss details of the compound structure and quality. One approach to have a more complete view of the analyzed compound is to use different separation principles like Electrophoresis and Chromatography. Due to its different separation modes (separations based on compound mobility or on adsorption/desorption characteristics) scientists can expect real different results for the same complex mixture analyzed. The seminar provides some examples of peptide mapping by comparing CE/MS separations to LC/MS separations.
Another way using orthogonal effects with the same separation principle is Two-dimensional liquid chromatography (2D-LC). Here different stationary phases and selectivity’s are used for complex mixtures to improve resolving power over conventional one-dimensional liquid chromatography. The seminar provides a basic overview on 2D-LC techniques and some separation examples of Biopharma relevant compounds.
Science Café Lunch Wednesday March 29, 12.15-13.15, room: Sorbonne 2, presented by Isogen Life Science
Whole Column- Imaged Capillary Iso-Electric Focusing for Protein Analysis
Dr. Martin Donker, (Isogen Life Science B.V., the Netherlands)
From unknown to having a charge based separation of your protein in only half a day is already common knowledge. But getting MS data from your charge based separation results is now available. The lunch session will be about the latest developments in whole column-imaged capillary IEF (WC-iCIEF).