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Originally published online as doi:10.2353/jmoldx.2007.060143 on May 24, 2007

Published online before print May 24, 2007
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Journal of Molecular Diagnostics 2007, Vol. 9, No. 3
Copyright © 2007 American Society for Investigative Pathology & Association for Molecular Pathology
DOI: 10.2353/jmoldx.2007.060143

Layered Peptide Array for Multiplex Immunohistochemistry

Gallya Gannot*, Michael A. Tangrea*, Heidi S. Erickson*, Peter A. Pinto{dagger}, Stephen M. Hewitt{ddagger}, Rodrigo F. Chuaqui*, John W. Gillespie§ and Michael R. Emmert-Buck*

From the Pathogenetics Unit, * Laboratory of Pathology and Urologic Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda; the Department of Urologic Oncology, {dagger} National Cancer Institute, National Institutes of Health, Bethesda; the Tissue Array Research Program, {ddagger} Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda; and Science Applications International Corporation, § National Cancer Institute, Bethesda, Maryland

Layered peptide array is a new methodology for multiplex molecular measurements from two-dimensional life science platforms. The technology can be used in several different configurations depending on the needs of the investigator. Described here is an indirect layered peptide array (iLPA) that is capable of measuring proteins on a solid surface, such as target antigens on a tissue section. A prototype iLPA system was developed and subsequently examined for reproducibility and specificity and then compared with standard immunohistochemistry. Semiquantitative, multiplex proteomic analysis of histological sections was achieved with up to 20 membranes. The experimental variability was 18%. Overall, the data suggest that iLPA technology will be a relatively simple and inexpensive method for molecular measurements from tissue sections.




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