Technical Report abstract
Nature Medicine 15, 566 - 571 (2009)
Published online: 12 April 2009 | doi:10.1038/nm.1903
Nanofluidic proteomic assay for serial analysis of oncoprotein activation in clinical specimens
Alice C Fan1, Debabrita Deb-Basu2, Mathias W Orban1, Jason R Gotlib3, Yasodha Natkunam4, Roger O'Neill2, Rose-Ann Padua5, Liwen Xu1, Daryl Taketa2, Amy E Shirer1, Shelly Beer1, Ada X Yee1, David W Voehringer2 & Dean W Felsher1
Abstract
Current methods of protein detection are insensitive to detecting subtle changes in oncoprotein activation that underlie key cancer signaling processes. The requirement for large numbers of cells precludes serial tumor sampling for assessing a response to therapeutics. Therefore, we have developed a nanofluidic proteomic immunoassay (NIA) to quantify total and low-abundance protein isoforms in nanoliter volumes. Our method can quantify amounts of MYC oncoprotein and B cell lymphoma protein-2 (BCL2) in Burkitt's and follicular lymphoma; identify changes in activation of extracellular signal–related kinases-1 (ERK1) and ERK2, mitogen-activated kinase-1 (MEK), signal transducer and activator of transcription protein-3 (STAT3) and STAT5, c-Jun N-terminal kinase (JNK) and caspase-3 in imatinib-treated chronic myelogeneous leukemia (CML) cells; measure an unanticipated change in the phosphorylation of an ERK2 isomer in individuals with CML who responded to imatinib; and detect a decrease in STAT3 and STAT5 phosphorylation in individuals with lymphoma who were treated with atorvastatin. Therefore, we have described a new and highly sensitive method for determining oncoprotein expression and phosphorylation in clinical specimens for the development of new therapeutics for cancer.
- Stanford University, School of Medicine, Division of Oncology, Departments of Medicine and Pathology, Stanford, California, USA.
- Cell Biosciences, Palo Alto, California, USA.
- Stanford University, School of Medicine, Division of Hematology, Department of Medicine, Stanford Cancer Center, Stanford, California, USA.
- Stanford University, School of Medicine, Department of Pathology, Stanford, California, USA.
- Institut Universitaire d'Hematologie, Institut National de la Santé et de la Recherche Médicale U718, Hopital Saint Louis, Paris, France.
Correspondence to: Dean W Felsher1 e-mail: dfelsher@stanford.edu
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