Edward S. Smith
Hoffmann-La Roche
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Publication
Featured researches published by Edward S. Smith.
PLOS ONE | 2012
Rajesh Patel; Alison Tsan; Rachel Tam; Rupal Desai; Nancy Schoenbrunner; Thomas W. Myers; Keith Bauer; Edward S. Smith; Rajiv Raja
Targeted anticancer therapies rely on the identification of patient subgroups most likely to respond to treatment. Predictive biomarkers play a key role in patient selection, while diagnostic and prognostic biomarkers expand our understanding of tumor biology, suggest treatment combinations, and facilitate discovery of novel drug targets. We have developed a high-throughput microfluidics method for mutation detection (MUT-MAP, mutation multi-analyte panel) based on TaqMan or allele-specific PCR (AS-PCR) assays. We analyzed a set of 71 mutations across six genes of therapeutic interest. The six-gene mutation panel was designed to detect the most common mutations in the EGFR, KRAS, PIK3CA, NRAS, BRAF, and AKT1 oncogenes. The DNA was preamplified using custom-designed primer sets before the TaqMan/AS-PCR assays were carried out using the Biomark microfluidics system (Fluidigm; South San Francisco, CA). A cross-reactivity analysis enabled the generation of a robust automated mutation-calling algorithm which was then validated in a series of 51 cell lines and 33 FFPE clinical samples. All detected mutations were confirmed by other means. Sample input titrations confirmed the assay sensitivity with as little as 2 ng gDNA, and demonstrated excellent inter- and intra-chip reproducibility. Parallel analysis of 92 clinical trial samples was carried out using 2–100 ng genomic DNA (gDNA), allowing the simultaneous detection of multiple mutations. DNA prepared from both fresh frozen and formalin-fixed, paraffin-embedded (FFPE) samples were used, and the analysis was routinely completed in 2–3 days: traditional assays require 0.5–1 µg high-quality DNA, and take significantly longer to analyze. This assay can detect a wide range of mutations in therapeutically relevant genes from very small amounts of sample DNA. As such, the mutation assay developed is a valuable tool for high-throughput biomarker discovery and validation in personalized medicine and cancer drug development.
Archive | 2011
Meike Eickhoff; Eberhard Russmann; Dirk Zimmermann; Andreas Wölfelschneider; Christopher Newhouse; Edward S. Smith; Sean Boyle
Archive | 2009
Keith Bauer; Ellen Fiss; David H. Gelfand; Edward S. Smith; Shawn Suko; Thomas W. Myers
CSH Protocols | 2006
Edward S. Smith; Alvin K. Li; Alice M. Wang; David H. Gelfand; Thomas W. Myers
Archive | 2017
Eickhoff Meike; Eberhard Russmann; Dirk Zimmermann; Andreas Woelfelschneider; Christopher Newhouse; Edward S. Smith; Sean Boyle
Archive | 2011
Meike Eickhoff; Eberhard Russmann; Dirk Zimmermann; Andreas Woelfelschneider; Christopher Newhouse; Edward S. Smith; Sean Boyle
Archive | 2011
Dirk Zimmermann; Meike Eickhoff; Eberhard Russmann; Andreas Wölfelschneider; Christopher Newhouse; Edward S. Smith; Sean Boyle
Archive | 2011
Meike Eickhoff; Eberhard Russmann; Dirk Zimmermann; Andreas Wölfelschneider; Christopher Newhouse; Edward S. Smith; Sean Boyle
Archive | 2011
Dirk Zimmermann; Meike Eickhoff; Eberhard Russmann; Andreas Wölfelschneider; Christopher Newhouse; Edward S. Smith; Sean Boyle
Archive | 2011
Dirk Zimmermann; Meike Eickhoff; Eberhard Russmann; Andreas Wölfelschneider; Christopher Newhouse; Edward S. Smith; Sean Boyle