LC-SWATH/MS Metabolomics Platform with Hyphenation of Extraction for the Analysis of Polar and Non-polar Metabolites in Biological Samples
Poster Oct 17, 2017
Michel Raetz 1, Renzo Picenoni 2, Guenter Boehm 2, Gérard Hopfgartner 1
Metabolomics studies are still challenging and technically demanding. Both sample preparation and data acquisition remain the bottlenecks in the measurements. On one hand, due to the large
chemical space of the analytes and their broad dynamic range that need to be dealt with. On the other hand, metabolomics still suffer from limited selectivity due to the structural similarities
within the low molecular weight compounds.
SWATH/MS opened new possibilities within Data Independent Aquisition (DIA) to gain selectivity for -omics studies as all fragments form all precursors in different isolation windows are recorded.
The hyphenated setup allows full automation from sample preparation over chromatography to MS data acquisition and feature annotation. The integrated dual LC configuration with three different
mobile phases enables a broad coverage of the metabolome. Additionally, the online SPE based lipid fractionation allows to equalize the dynamic range of the abundance of lipid classes.
Multiplexing cell-based assays is possible using 3D culture models that are larger and more complex than monolayers
Real-time detection methods to measure live or dead cells provide much flexibility for multiplexing
All multiplexed assay combinations should be verified using appropriate controls for each 3D cell culture model.
Basic fibroblast growth factor (bFGF) is widely used in vitro for the maintenance and stimulation of a variety of cells. However, use of native bFGF in cell biology is limited by the fact that bFGF rapidly degrades at physiological temperatures. We have addressed this problem with an engineered form of bFGF, named Heat Stable bFGF (HS bFGF), which is stable at 37 degrees Celsius.READ MORE