omation mbers - Society for Laboratory Automation and Screening

3:00 pm Tuesday, February 3 Proteomics – Arrays Room B1
Brian Haab
Van Andel Research Institute
333 Bostwick
Grand Rapids, Michigan 49503
brian.haab@vai.org
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Co-Author(s)
Heping Zhou, Mark Schotanus, Randall Brand
Evanston Northwestern Hospital
Jorge Marrero
University of Michigan Medical School
Deborah Dillon, Jose Costa, Paul Lizardi
Yale School of Medicine
High Sensitivity Multiplexed Serum Protein Analysis Using Antibody Microarrays and
Rolling Circle Amplification
Antibody microarrays enable highly multiplexed and rapid protein measurements in low sample volumes. The
profiling of proteins in sera and other bodily fluids using this tool should offer new opportunities for biomarker
discovery and insights into disease biology. Robotically spotted microarrays of antibodies and proteins were used
to measure the relative abundances of multiple proteins in serum samples from prostate cancer and pancreatic
cancer patients and controls. Serum proteins that had been coupled to either a fluorescent tag (e.g., Cy3) or a
hapten (e.g., biotin) were incubated on the microarrays, and specific proteins bound to the immobilized molecules
on the microarrays through specific interactions. After washing away unbound proteins, bound proteins were
detected using the fluorescent tag or amplified signal (using rolling circle amplification, RCA) from the haptenlabeled
proteins. RCA significantly enhanced detection sensitivity while maintaining the accuracy and precision
of the measurements. Measurements of dozens of proteins in sera from cancer patients and controls revealed
significant differences in protein abundances between the two sample groups. The biological significance and
potential clinical usefulness of the observed protein alterations in the sera of cancer patients will be discussed.
3:30 pm Tuesday, February 3 Proteomics – Arrays Room B1
Paul Predki
Protometrix, Inc.
688 E. Main Street
Branford, Connecticut 06405
paul.predki@protometrix.com
Application of Functional Protein Microarrays to Kinase Drug R&D
The ability to use protein microarrays to analyze protein function and interactions with a wide variety of molecules,
including proteins, lipids, DNA, substrates, antibodies and drugs, has only recently been realized. Protometrix has
industrialized and validated the complete process required for the development of proteome and sub-proteome
microarrays for these applications. A yeast proteome array (the yeast ProtoArray) has already been completed,
and a variety of human sub-proteome arrays are under development. Among these are human kinase arrays,
which we have now validated for both interaction and activity assays. These simple protocols will provide powerful
new capabilities when applied to the drug R&D process. Examples of these applications include kinase substrate
identification, interaction and pathway mapping, antibody validation, inhibitor activity profiling and proteome-scale
determination of inhibitor binding specificity. Recent results in each of these areas will be discussed.