Layered Peptide Arrays: High-Throughput Antibody Screening of Clinical Samples

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...

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Published inThe Journal of molecular diagnostics : JMD Vol. 7; no. 4; pp. 427 - 436
Main Authors Gannot, Gallya, Tangrea, Michael A, Gillespie, John W, Erickson, Heidi S, Wallis, Benjamin S, Leakan, Rose Anne, Knezevic, Vladimir, Hartmann, Dan P, Chuaqui, Rodrigo F, Emmert-Buck, Michael R
Format Journal Article
LanguageEnglish
Published United States ASIP 01.10.2005
American Society for Investigative Pathology
Subjects
Antibodies - analysis
Antibodies - immunology
Cluster Analysis
Humans
Mass Screening - instrumentation
Mass Screening - methods
Peptides - chemistry
Peptides - immunology
Protein Array Analysis - instrumentation
Protein Array Analysis - methods
Regular
Saliva - immunology
Sjogren's Syndrome - blood
Sjogren's Syndrome - diagnosis
Sjogren's Syndrome - immunology
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Summary: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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Address reprint requests to Michael R. Emmert-Buck, Pathogenetics Unit, Advanced Technology Center, Laboratory of Pathology and Urologic Oncology Branch, Center for Cancer Research, National Cancer Institute, 8717 Grovemont Circle, Bethesda, MD 20892-4605. E-mail: mbuck@helix.nih.gov.
ISSN:1525-1578
1943-7811
DOI:10.1016/S1525-1578(10)60573-9