{"doi":"10.3390/antib14040097","title":"A Novel FLI1 Monoclonal Antibody Which Recognizes EWS::FLI1 with High Affinity Is Useful for Detecting Ewing Sarcoma","abstract":"BACKGROUND: Ewing sarcoma (ES) is a rare tumor that affects children, adolescents, and young adults. ES is associated with high morbidity in all patients and high mortality for those who present with metastatic disease. A chromosomal translocation, either t(11;22)(q24;p12) or t(21;22)(q22;q12) leads to the fusion oncoproteins EWS::FLI1 or EWS::ERG in 95% of ES patients. We recognized a critical need for a stably sourced high-affinity antibody that recognizes EWS::FLI1 with maximal specificity. Understanding EWS::FLI1 protein complexes is a pivotal gap in ES knowledge that necessitates the development of antibodies capable of identifying native proteins in solution. Further, variable epitope sequencing of a monoclonal antibody enables the construction of degraders and nanobody identifiers. METHODS: Monoclonal antibodies were produced following informed peptide synthesis, injection, and hybridoma creation. Hybridoma antibodies were validated for specificity and function. RESULTS: Our results indicate that the FLI1 1.2 monoclonal antibody, which recognizes the EWS::FLI1 fusion oncoprotein, can be reliably applied to multiple molecular biology applications like immunoblot, immunoprecipitation, immunofluorescence, and immunohistochemistry. This FLI1 1.2 monoclonal antibody has a high affinity of 0.3 nM KD to EWS::FLI1. In terms of specificity, this antibody is highly specific to EWS::FLI1 and some cross reactivity with ERG. CONCLUSIONS: This reagent will provide the research community with valuable tools for further biochemical and genomic interrogation of the oncogenic activity of EWS::FLI1 in ES.","journal":"Antibodies","year":2025,"id":548416,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9633,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1442102,"name":"Olivia O. Lansinger","orcid":null,"position":1,"is_corresponding":false},{"id":805667,"name":"David V. Allegakoen","orcid":"0000-0002-0261-2962","position":2,"is_corresponding":false},{"id":1442103,"name":"Emma J. W. McGuire","orcid":null,"position":3,"is_corresponding":false},{"id":1442104,"name":"Ashley R. Gaffey","orcid":null,"position":4,"is_corresponding":false},{"id":1442105,"name":"Jeff R. Petro","orcid":null,"position":5,"is_corresponding":false},{"id":397586,"name":"Purushottam B. Tiwari","orcid":"0000-0002-3140-0723","position":6,"is_corresponding":false},{"id":1442106,"name":"Quinn Tufiño","orcid":null,"position":7,"is_corresponding":false},{"id":397588,"name":"Aykut Üren","orcid":"0000-0002-2033-1216","position":8,"is_corresponding":false},{"id":468196,"name":"Jeffrey A. Toretsky","orcid":"0000-0003-3619-2879","position":9,"is_corresponding":false},{"id":955059,"name":"Saravana P. Selvanathan","orcid":"0000-0002-6990-5188","position":0,"is_corresponding":true}],"reference_count":37,"raw_metadata":null,"created_at":"2026-07-19T02:53:58.530220Z","pmid":"41283512","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":[]}