Development and optimization of a procedure for extracting proteins from fixed archival tissues and assessment of its potential for biomarker discovery in human and veterinary medicine
Poster Dec 18, 2013
Alessandro Tanca, Daniela Pagnozzi, Sergio Uzzau, Maria Filippa Addis
Fixed tissue repositories are an ideal information mine for biomarker discovery, with their vast collections of diseased tissues and associated clinical records describing diagnosis, prognosis, therapy, and outcome. Unfortunately, fixation hampers protein extraction, posing considerable limitations to proteomic investigations. In this scenario, and building on the existing literature, several methods were combined and refined to enable extraction of high-quality, full-length proteins from the fixed specimens, leading to the definition of an optimized protocol with unprecedented performances. In order to test its ability to provide meaningful results in terms of differential disease traits and enable biomarker discovery, the optimized method was applied to sample repositories stored in pathology departments. Specifically, diseased human, canine and sheep tissues were retrieved and subjected to the optimized extraction procedure, followed by established proteomic pipelines consisting of electrophoresis, image analysis, mass spectrometry, and data processing by ontology and pathway analysis. The method enabled gel-based proteomic investigations of archival tissues, performing better than commercially available systems. Differential proteins were successfully identified in all cases, and these were clearly related to the disease biology. In conclusion, biomarker discovery strategies based on proteomic investigations can be successfully performed on fixed archival tissues by means of this optimized extraction protocol.
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.
A New Tool for the Automated Sample Preparation of Whole Blood Samples by LC-MS using a Commercial AutosamplerPoster
Automated sample preparation reduces the costs per sample and minimizes sample handling errors. Usually expensive and highly specialized pipetting robots are used. However, most of these systems are not designed with a direct interface for LC-MS applications. In addition common pipetting systems are not optimized for smaller scale sample series. Here we present a new tool for liquid handling of whole blood samples and direct sample injection.READ MORE
Fully automated sample preparation for the determination of plasticizers in PVC from food contact materials and toysPoster
The poster outlines a comprehensive, fully automated analysis of plasticizers in toys and food stuff.READ MORE