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JMD 2005, Vol. 7, No. 4
Copyright © 2005 American Society for Investigative Pathology & Association for Molecular Pathology

Layered Peptide Arrays

High-Throughput Antibody Screening of Clinical Samples

Gallya Gannot^*, Michael A. Tangrea^*, John W. Gillespie^*, Heidi S. Erickson^*, Benjamin S. Wallis^*, Rose Anne Leakan, Vladimir Knezevic, Dan P. Hartmann, Rodrigo F. Chuaqui^* 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, Maryland; the Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland; 20/20 GeneSystems, Rockville, Maryland; and Georgetown University, Washington, District of Columbia

High-throughput methods to detect and quantify antibodies in sera and other patient specimens have use for many clinical and laboratory studies, including those associated with cancer detection, microbial exposures, and autoimmune diseases. We developed a new technique, termed layered peptide array (LPA), to serve as a screening tool to detect antibodies in a highly multiplexed format. We demonstrate here that a prototype LPA was capable of producing approximately 5000 measurements per experiment and appeared to be scalable to higher throughput levels. Sera and saliva from Sjögren’s syndrome patients served as a test set to examine antibody titers in clinical samples. The LPA platform exhibited both a high sensitivity (100%) and high specificity (94%) for correctly identifying SSB antigen-positive samples. The multiplex capability of the platform was also confirmed when serum and saliva samples were analyzed for antibody reactivity to several peptides, including Sjögren’s syndrome antigens A and B. The data indicate that LPA analysis will be a useful method for a number of screening applications.

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				G. GANNOT, M. A TANGREA, R. F CHUAQUI, J. W GILLESPIE, and M. R EMMERT-BUCK Layered Peptide Arrays: A Diverse Technique for Antibody Screening of Clinical Samples Ann. N.Y. Acad. Sci., March 1, 2007; 1098(1): 451 - 453. [Abstract] [Full Text] [PDF]