{"doi":"10.1093/braincomms/fcaa059","title":"High-resolution epitope mapping of anti-Hu and anti-Yo autoimmunity by programmable phage display","abstract":"Abstract 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.","journal":"Brain Communications","year":2020,"id":59402,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":67,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.8909,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":308641,"name":"Caleigh Mandel‐Brehm","orcid":"0000-0001-7764-6529","position":1,"is_corresponding":false},{"id":308639,"name":"Sara E. Vazquez","orcid":"0000-0002-0601-7001","position":2,"is_corresponding":false},{"id":312357,"name":"Jamin Liu","orcid":"0000-0001-6386-3709","position":3,"is_corresponding":false},{"id":312358,"name":"Audrey V. Parent","orcid":"0000-0003-4506-6268","position":4,"is_corresponding":false},{"id":235418,"name":"Mark S. Anderson","orcid":"0000-0002-3093-4758","position":5,"is_corresponding":false},{"id":313261,"name":"Travis Kassimatis","orcid":null,"position":6,"is_corresponding":false},{"id":312359,"name":"Αναστασία Ζεκερίδου","orcid":"0000-0002-1960-9065","position":7,"is_corresponding":false},{"id":246852,"name":"Stephen L. Hauser","orcid":"0000-0002-4932-4001","position":8,"is_corresponding":false},{"id":231254,"name":"Sean J. Pittock","orcid":"0000-0002-6140-5584","position":9,"is_corresponding":false},{"id":108693,"name":"Eric D. Chow","orcid":"0000-0001-8079-918X","position":10,"is_corresponding":false},{"id":109296,"name":"Michael R. Wilson","orcid":"0000-0002-8705-5084","position":11,"is_corresponding":false},{"id":107235,"name":"Joseph L. DeRisi","orcid":"0000-0002-4611-9205","position":12,"is_corresponding":false},{"id":312356,"name":"Brian D. O’Donovan","orcid":"0000-0002-2901-7530","position":0,"is_corresponding":true}],"reference_count":48,"raw_metadata":null,"created_at":"2026-07-18T21:07:59.189921Z","pmid":"32954318","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}