Summary
Analysis of the genome provides important information about the somatic genetic changes existing in the tissue; however, it is the proteins that do the work of the cell. Diseases such as cancer are caused by derangements in cellular protein molecular networks and cell signaling pathways. These pathways contain a large and growing collection drug targets, governing cellular survival, proliferation, invasion, and cell death. The clinical utility of reverse-phase protein microarrays (RPPA), a new technology invented in our laboratory, lies in its ability to generate a functional map of known cell signaling networks or pathways for an individual patient obtained directly from a biopsy specimen. Coupled with laser capture microdissection (LCM), the RPPA platform, the entire cellular proteome is immobilized on a substratum with subsequent immunodetection of the total levels and phosphorylated, or activated, state of cell signaling proteins. The results of which pathways are “in use” can then be correlated with biological and clinical information and serve as both a diagnostic and a therapeutic guide, thus providing a “theranostic” endpoint.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Liotta L. A., Espina V., Mehta A. I., (2003) Protein microarrays: Meeting analytical challenges for clinical applications. Cancer Cell 3(4), 317–25.
Haab B. B., Dunham M. J., and Brown P. O. (2001) Protein microarrays for highly parallel detection and quantitation of specific proteins and antibodies in complex solutions. Genome Biol 2(2), RESEARCH0004.
Macbeath G. and Schreiber S. L. (2000) Printing proteins as microarrays for high-throughput function determination. Science 289(5485), 1760–3.
Macbeath G. (2002) Protein microarrays and proteomics. Nat Genet 32, 526–32.
Paweletz C. P., Charboneau L., Bichsel V. E., (2001) Reverse phase protein microarrays which capture disease progression show activation of pro-survival pathways at the cancer invasion front. Oncogene 20(16), 1981–9.
Zhu H. and Snyder M. (2003) Protein chip technology. Curr Opin Chem Biol 7(1), 55–63.
Wilson D. S. and Nock S. (2003) Recent developments in protein microarray technology. Angew Chem Int Ed Engl 42(5), 494–500.
Templin M. F., Stoll D., Schrenk M. (2002) Protein microarray technology. Trends Biotechnol 20(4), 160–6.
Schaeferling M., Schiller S., Paul H., (2002) Application of self-assembly techniques in the design of biocompatible protein microarray surfaces. Electrophoresis 23(18), 3097–105.
Weng S., Gu K., Hammond P. W.. (2002) Generating addressable protein microarrays with PROfusion covalent mRNA-protein fusion technology. Proteomics 2(1), 48–57.
Petach H. and Gold L. (2002) Dimensionality is the issue: use of photoaptamers in protein microarrays. Curr Opin Biotechnol 13(309–314.
Lal S. P., Christopherson R. I., and Dos Remedios C. G. (2002) Antibody arrays: an embryonic but rapidly growing technology. Drug Discov Today 7(18 Suppl), S143–9.
Humphery-Smith I., Wischerhoff E., and Hashimoto R. (2002) Protein arrays for assessment of target selectivity. Drug Discov World 4(1), 17–27.
Bobrow M. N., Harris T. D., Shaughnessy K. J., and Litt G. J. (1989) Catalyzed reporter deposition, a novel method of signal amplification. Application to immunoassays. J Immunol Methods 125(1–2), 279–85.
Bobrow M. N., Shaughnessy K. J., and Litt G. J. (1991) Catalyzed reporter deposition, a novel method of signal amplification. II. Application to membrane immunoassays. J Immunol Methods 137(1), 103–12.
Hunyady B., Krempels K., Harta G., and Mezey E. (1996) Immunohistochemical signal amplification by catalyzed reporter deposition and its application in double immunostaining. J Histochem Cytochem 44(12), 1353–62.
King G., Payne S., Walker F., and Murray G. I. (1997) A highly sensitive detection method for immunohistochemistry using biotinylated tyramine. J Pathol 183(2), 237–41.
Petricoin E., Wulfkuhle J., Espina V., and Liotta L. A. (2004) Clinical proteomics: revolutionizing disease detection and patient tailoring therapy. J Proteome Res 3(2), 209–17.
Grubb R. L., Calvert V. S., Wulkuhle J. D., (2003) Signal pathway profiling of prostate cancer using reverse phase protein arrays. Proteomics 3(11), 2142–6.
Wulfkuhle J. D., Aquino J. A., Calvert V. S., (2003) Signal pathway profiling of ovarian cancer from human tissue specimens using reverse-phase protein microarrays. Proteomics 3(11), 2085–90.
Liotta L. A., Kohn E. C., and Petricoin E. F. (2001) Clinical proteomics: personalized molecular medicine. JAMA 286(18), 2211–4.
Petricoin E. F., Zoon K. C., Kohn E. C., Barrett J. C., and Liotta L. A. (2002) Clinical proteomics: translating benchside promise into bedside reality. Nat Rev Drug Discov 1(9), 683–95.
Wulfkuhle J. D., Edmiston K. H., Liotta L. A., and Petricoin E. F. (2006) Technology Insight: pharmacoproteomics for cancer-promises of patient-tailored medicine using protein microarrays. Nat Clin Pract Oncol 3(5):256–68.
Berggren K., Steinberg T. H., Lauber W. M., (1999) A luminescent ruthenium complex for ultrasensitive detection of proteins immobilized on membrane supports. Anal Biochem 276(2), 129–43.
Tonkinson J. L. and Stillman B. A. (2002) Nitrocellulose: a tried and true polymer finds utility as a post-genomic substrate. Front Biosci 7, c1–12.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Espina, V., Liotta, L., Petricoin, E. (2009). Reverse-Phase Protein Microarrays for Theranostics and Patient Tailored Therapy. In: Tainsky, M. (eds) Tumor Biomarker Discovery. Methods in Molecular Biology, vol 520. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-811-9_7
Download citation
DOI: https://doi.org/10.1007/978-1-60327-811-9_7
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-810-2
Online ISBN: 978-1-60327-811-9
eBook Packages: Springer Protocols