Abstract
Immune responses targeting self-proteins (autoantigens) can lead to a variety of autoimmune diseases. Identification of these antigens is important for both diagnostic and therapeutic reasons. However, current approaches to characterize autoantigens have, in most cases, met only with limited success. Here we present a synthetic representation of the complete human proteome, the T7 peptidome phage display library (T7-Pep), and demonstrate its application to autoantigen discovery. T7-Pep is composed of >413,000 36-residue, overlapping peptides that cover all open reading frames in the human genome, and can be analyzed using high-throughput DNA sequencing. We developed a phage immunoprecipitation sequencing (PhIP-Seq) methodology to identify known and previously unreported autoantibodies contained in the spinal fluid of three individuals with paraneoplastic neurological syndromes. We also show how T7-Pep can be used more generally to identify peptide-protein interactions, suggesting the broader utility of our approach for proteomic research.
Publication types
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Research Support, American Recovery and Reinvestment Act
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Amino Acid Sequence
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Antigens, Neoplasm / immunology
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Autoantibodies / immunology
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Autoantibodies / metabolism
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Autoantigens / genetics
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Autoantigens / immunology*
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Autoantigens / isolation & purification*
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Autoimmune Diseases / genetics
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Autoimmune Diseases / immunology
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Bacteriophage T7 / metabolism
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Carcinoma, Non-Small-Cell Lung / immunology
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Cloning, Molecular
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Female
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Gene Library
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Genome, Human
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High-Throughput Nucleotide Sequencing
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Humans
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Immunoprecipitation
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Middle Aged
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Molecular Sequence Data
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Nerve Tissue Proteins / immunology
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Neuro-Oncological Ventral Antigen
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Oligonucleotide Array Sequence Analysis
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Open Reading Frames / immunology
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Paraneoplastic Syndromes, Nervous System / immunology
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Peptide Library*
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Proteome / genetics*
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Proteomics / methods*
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RNA-Binding Proteins / immunology
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Sequence Analysis, RNA
Substances
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Antigens, Neoplasm
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Autoantibodies
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Autoantigens
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Nerve Tissue Proteins
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Neuro-Oncological Ventral Antigen
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Peptide Library
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Proteome
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RNA-Binding Proteins