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High-resolution epitope mapping of anti-Hu and anti-Yo autoimmunity by programmable phage display.

  • O'Donovan, Brian1
  • Mandel-Brehm, Caleigh1
  • Vazquez, Sara E1
  • Liu, Jamin1, 2
  • Parent, Audrey V3
  • Anderson, Mark S3
  • Kassimatis, Travis1
  • Zekeridou, Anastasia4, 5
  • Hauser, Stephen L6, 7
  • Pittock, Sean J4, 5
  • Chow, Eric1
  • Wilson, Michael R6, 7
  • DeRisi, Joseph L1, 8
  • 1 Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94158, USA.
  • 2 UC Berkeley-UCSF Graduate Program in Bioengineering, University of California, Berkeley, Berkeley, CA 94158, USA.
  • 3 Department of Medicine, Diabetes Center, University of California, San Francisco, San Francisco, CA 94143, USA.
  • 4 Department of Neurology, Mayo Clinic, Rochester, MN 55902, USA.
  • 5 Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN 55902, USA.
  • 6 Weill Institute for Neurosciences, University of California, San Francisco, San Francisco, CA 94158, USA.
  • 7 Department of Neurology, University of California, San Francisco, San Francisco, CA 94158, USA.
  • 8 Chan Zuckerberg Biohub, University of California, San Francisco, San Francisco, CA 94158, USA.
Published Article
Brain communications
Publication Date
Jan 01, 2020
DOI: 10.1093/braincomms/fcaa059
PMID: 32954318


Paraneoplastic neurological disorders are immune-mediated diseases understood to manifest as part of a misdirected anti-tumor immune response. Paraneoplastic neurological disorder-associated autoantibodies can assist with diagnosis and enhance our understanding of tumor-associated immune processes. We designed a comprehensive library of 49-amino-acid overlapping peptides spanning the entire human proteome, including all splicing isoforms and computationally predicted coding regions. Using this library, we optimized a phage immunoprecipitation and sequencing protocol with multiple rounds of enrichment to create high-resolution epitope profiles in serum and cerebrospinal fluid (CSF) samples from patients suffering from two common paraneoplastic neurological disorders, the anti-Yo (n = 36 patients) and anti-Hu (n = 44 patients) syndromes. All (100%) anti-Yo patient samples yielded enrichment of peptides from the canonical anti-Yo (CDR2 and CDR2L) antigens, while 38% of anti-Hu patients enriched peptides deriving from the nELAVL (neuronal embryonic lethal abnormal vision like) family of proteins, the anti-Hu autoantigenic target. Among the anti-Hu patient samples that were positive for nELAVL, we noted a restricted region of immunoreactivity. To achieve single amino acid resolution, we designed a novel deep mutational scanning phage library encoding all possible single-point mutants targeting the reactive nELAVL region. This analysis revealed a distinct preference for the degenerate motif, RLDxLL, shared by ELAVL2, 3 and 4. Lastly, phage immunoprecipitation sequencing identified several known autoantigens in these same patient samples, including peptides deriving from the cancer-associated antigens ZIC and SOX families of transcription factors. Overall, this optimized phage immunoprecipitation sequencing library and protocol yielded the high-resolution epitope mapping of the autoantigens targeted in anti-Yo and anti-Hu encephalitis patients to date. The results presented here further demonstrate the utility and high-resolution capability of phage immunoprecipitation sequencing for both basic science and clinical applications and for better understanding the antigenic targets and triggers of paraneoplastic neurological disorders. © The Author(s) (2020). Published by Oxford University Press on behalf of the Guarantors of Brain.

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