Automated Sample Preparation
On the basis of the liquid handling workstation Tecan Freedom Evo 150, we have established a 3-in-1 robotic system for fully automated processing of 2D gel-derived gel plugs, including in-gel digestion, extraction of tryptic peptides, and MALDI target spotting. The key component of the process is our stack-type digestion device, which has been licensed to Tecan (pdf brochure). We prepare the samples for MALDI-MS according to the thin-layer affinity method originally described by Gobom et al. (Anal Chem 2001). For this purpose, Bruker AnchorChip targets are automatically coated with a microcrystalline layer of CHCA matrix using our motor-operated matrix application device. Recently, we have developed another digestion device tailored to the purpose of processing larger gel pieces of variable sizes. This system is routinely used for protein identification from single 1D gel-derived gel bands or – in combination with a gel punching device – from entire 1D gel lanes.
For further information, please refer to our original publications on automation advances in gel-based proteomics:
Jahn, O.; Hesse, D.; Reinelt, M.; Kratzin, H. D. Technical innovations for the automated identification of gel-separated proteins by MALDI-TOF mass spectrometry. Analytical and Bioanalytical Chemistry 2006; 386:92-103. Abstract.
Schmidt, C.; Hesse, D.; Raabe, M.; Urlaub, H.; Jahn, O. An automated in-gel digestion / iTRAQ-labeling workflow for robust quantification of gel-separated proteins. Proteomics 2013; 13:1417-22. Abstract.