Automated protein digestion workflows for MS-based proteomics applications
Poster Jun 18, 2014
Gunnar Dittmar1, Oliver Popp1, Guenter Boehm2, Andreas Bruchmann3
Mass spectrometry (MS) based bottom-up proteomics is built upon large scale identification of peptides, and depends on proteins being efficiently converted to peptides by a protease of known specificity. The most common preparation methods are digestion in solution (ISD) or digestion of Proteins separated on an SDS PAGE gel, in-gel digestion (IGD). Both methods consist of a lengthy sequence of washing and chemical modification steps. To increase throughput and reproducibility, automation of these processes is highly desired. Contrary to other „omic“-applications, proteomics analysis by LC-MS/MS remains time-intensive, making the measurement the ratedetermining step in the pipeline. Thus the preparation of samples does not require a high-but rather a medium-throughput setup.
The normal benchtop methods for the IGD and the ISD were adapted to make automation in a robotic setup possible. In addition, we modified a standard PAL robot setup with a vacuum chamber that can be controlled by the robot's software and facilitates the removal of large volumes of washing solvents required by IGD leading to enhanced sensitivity.
Proteomics and Substrate Based MS Imaging of Xenobiotic Metabolising Enzymes in Ex Vivo Human Skin and a Human Living Skin Equivalent ModelPoster
Untargeted proteomics analysis showed that human skin and a commercially available living skin equivalent model exhibit a similar distribution of xenobiotic metabolising enzymes. A new technique, substrate based mass spectrometry imaging (SB-MSI) was developed during this study.READ MORE
Development and Assessment of Non-Infected and Infected Skin Models Using MALDI-MSIPoster
Infections have detrimental effects on wound healing leading to chronic wound formation. The use of labskin,a living skin model, in combination with mass spectrometry imaging, provides a good model for assessing wound healing factors.READ MORE
Overcoming the Challenges of Analysing Ionic Polar Pesticides in FoodPoster
Here, the novel application of the Waters’ Torus DEA column is presented, showing the improved performance of a UPLC-MS/MS method for the underivatised
analysis of glyphosate and a selection of other anionic pesticides.