{"doi":"10.1126/scitranslmed.abn0402","title":"Ex silico engineering of cystine-dense peptides yielding a potent bispecific T cell engager","abstract":"Cystine-dense peptides (CDPs) are a miniprotein class that can drug difficult targets with high affinity and low immunogenicity. Tools for their design, however, are not as developed as those for small-molecule and antibody drugs. CDPs have diverse taxonomic origins, but structural characterization is lacking. Here, we adapted Iterative Threading ASSEmbly Refinement (I-TASSER) and Rosetta protein modeling software for structural prediction of 4298 CDP scaffolds and performed in silico prescreening for CDP binders to targets of interest. Mammalian display screening of a library of docking-enriched, methionine and tyrosine scanned (DEMYS) CDPs against PD-L1 yielded binders from four distinct CDP scaffolds. One was affinity-matured, and cocrystallography yielded a high-affinity ( K D = 202 pM) PD-L1–binding CDP that competes with PD-1 for PD-L1 binding. Its subsequent incorporation into a CD3-binding bispecific T cell engager produced a molecule with pM-range in vitro T cell killing potency and which substantially extends survival in two different xenograft tumor-bearing mouse models. Both in vitro and in vivo, the CDP-incorporating bispecific molecule outperformed a comparator antibody-based molecule. This CDP modeling and DEMYS technique can accelerate CDP therapeutic development.","journal":"Science Translational Medicine","year":2022,"id":272721,"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":11,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9537,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":475518,"name":"Emily J. Girard","orcid":"0000-0003-1857-0194","position":1,"is_corresponding":false},{"id":475519,"name":"Gregory P. Sevilla","orcid":"0000-0002-6590-9413","position":2,"is_corresponding":false},{"id":720935,"name":"Mi‐Youn Brusniak","orcid":"0000-0003-3950-1614","position":3,"is_corresponding":false},{"id":475520,"name":"Peter B. Rupert","orcid":"0000-0003-1011-0302","position":4,"is_corresponding":false},{"id":476315,"name":"Della Friend","orcid":null,"position":5,"is_corresponding":false},{"id":245713,"name":"Mesfin Gewe","orcid":"0000-0001-9587-7283","position":6,"is_corresponding":false},{"id":939274,"name":"Midori Clarke","orcid":"0000-0002-2402-108X","position":7,"is_corresponding":false},{"id":939717,"name":"Ida Lin","orcid":null,"position":8,"is_corresponding":false},{"id":475524,"name":"Raymond Ruff","orcid":"0000-0002-7189-1221","position":9,"is_corresponding":false},{"id":475521,"name":"Fiona Pakiam","orcid":"0000-0001-6292-1137","position":10,"is_corresponding":false},{"id":234630,"name":"Tinh-Doan Phi","orcid":null,"position":11,"is_corresponding":false},{"id":704218,"name":"Ashok D. Bandaranayake","orcid":null,"position":12,"is_corresponding":false},{"id":245717,"name":"Colin Correnti","orcid":"0000-0003-3710-9619","position":13,"is_corresponding":false},{"id":475527,"name":"Andrew J. Mhyre","orcid":"0000-0002-9206-3150","position":14,"is_corresponding":false},{"id":939275,"name":"Natalie W. Nairn","orcid":"0000-0003-1293-345X","position":15,"is_corresponding":false},{"id":475528,"name":"Roland K. Strong","orcid":"0000-0002-1338-2189","position":16,"is_corresponding":false},{"id":30180,"name":"James M. Olson","orcid":"0000-0001-5990-6534","position":17,"is_corresponding":false},{"id":475517,"name":"Zachary R. Crook","orcid":"0000-0002-1142-9748","position":0,"is_corresponding":true}],"reference_count":73,"raw_metadata":null,"created_at":"2026-07-19T00:27:47.926496Z","pmid":"35584229","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":[]}