Scientists at the NMI, the University of Tübingen, the Technical University of Munich and the University of Düsseldorf have established a novel approach for highly parallelized analyses of hundreds of proteins and post-translational protein modifications. The innovative methodology is based on a combination of classical Western blotting - the gold standard in protein detection - and the multiplexing power of Luminex® bead technology. This currently allows detection of up to 600 total proteins and protein modifications from a sample of cultured cells or in vivo-derived tissue, based on an ever-growing list of >1,000 validated high-quality antibodies. DigiWest® thus increases the power of a Western blot by a factor of 300, while minimizing the amount of sample required, enabling quantification from valuable, limited samples.
After successful technical benchmarking of the DigiWest® method, it was applied in two case studies to elucidate mechanisms of resistance to the anticancer drug Lapatinib in a cell-based experiment and to classify different forms of breast carcinoma from primary tumor sections. In both cases, they identified a number of promising candidate marker proteins, some of which were phosphorylated, convincingly demonstrating the superiority of protein profiling compared to classical DNA- and RNA-based biomarker discovery.
Markus Templin, Ph.D., head of assay development at the NMI and senior author of the publication, commented, "After a decade of genomic research, scientists are yearning for new technologies to analyze the complexity of cellular signal transduction at the level of protein activation, which has too often been the real shortcoming of genomics. Our DigiWest® methodology overcomes this limitation and provides a powerful and versatile proteomics-scale platform for a variety of applications, including lead compound profiling, pathway mapping, biomarker research, and precision medicine development."
NMI TT Pharmaservices, the contract research division of the NMI, has established dedicated laboratory capabilities to offer fully customizable DigiWest® studies on a fee-for-service basis to the industrial and academic life science community. For more information, visit the DigiWest® website: www.digiwest.de
The full text version of the DigiWest® publication can be accessed via the Nature Communications website: www.nature.com/ncomms/2016/160923/ncomms12852/full/ncomms1
