Application of a synthetic human proteome to autoantigen discovery through PhIP-Seq

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Larman, H. Benjamin
Zhao, Zhenming
Laserson, Uri
Li, Mamie Z.
Ciccia, Alberto
Gakidis, M. Angelica Martinez
Kesari, Santosh
LeProust, Emily M.
Solimini, Nicole L.
Note: Order does not necessarily reflect citation order of authors.
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https://doi.org/10.1038/nbt.1856Metadata
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Larman, H. B., Z. Zhao, U. Laserson, M. Z. Li, A. Ciccia, M. A. M. Gakidis, G. M. Church, et al. 2011. “Application of a synthetic human proteome to autoantigen discovery through PhIP-Seq.” Nature biotechnology 29 (6): 535-541. doi:10.1038/nbt.1856. http://dx.doi.org/10.1038/nbt.1856.Abstract
In this study, we improve on current autoantigen discovery approaches by creating a synthetic representation of the complete human proteome, the T7 “peptidome” phage display library (T7-Pep), and use it to profile the autoantibody repertoires of individual patients. We provide methods for 1) designing and cloning large libraries of DNA microarray-derived oligonucleotides encoding peptides for display on bacteriophage, and 2) analysis of the peptide libraries using high throughput DNA sequencing. We applied phage immunoprecipitation sequencing (PhIP-Seq) to identify both known and novel autoantibodies contained in the spinal fluid of three patients with paraneoplastic neurological syndromes. We also show how our approach can be used more generally to identify peptide-protein interactions and point toward ways in which this technology will be further developed in the future. We envision that PhIP-Seq can become an important new tool in autoantibody analysis, as well as proteomic research in general.Other Sources
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4169279/pdf/Terms of Use
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