{"doi":"10.1073/pnas.1920662117","title":"Structure-guided engineering of the affinity and specificity of CARs against Tn-glycopeptides","abstract":"-acetylgalactosamine (GalNAc, known as Tn antigen) rather than the normally extended carbohydrate. Previously, we used the scFv fragment of antibody 237 as a chimeric antigen receptor (CAR) to mediate recognition of mouse tumor cells that bear its cognate Tn-glycopeptide epitope in podoplanin, also called OTS8. Guided by the structure of the 237 Fab:Tn-OTS8-glycopeptide complex, here we conducted a deep mutational scan showing that residues flanking the Tn-glycan contributed significant binding energy to the interaction. Design of 237-scFv libraries in the yeast display system allowed us to isolate scFv variants with higher affinity for Tn-OTS8. Selection with a noncognate human antigen, Tn-MUC1, yielded scFv variants that were broadly reactive with multiple Tn-glycoproteins. When configured as CARs, engineered T cells expressing these scFv variants showed improved activity against mouse and human cancer cell lines defective in O-linked glycosylation. This strategy provides CARs with Tn-peptide specificities, all based on a single scFv scaffold, that allows the same CAR to be tested for toxicity in mice and efficacy against mouse and human tumors.","journal":"Proceedings of the National Academy of Sciences","year":2020,"id":91790,"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":44,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9499,"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":459419,"name":"Venkata V.V.R. Marada","orcid":"0000-0003-1142-7854","position":1,"is_corresponding":false},{"id":459420,"name":"Qi Cai","orcid":"0000-0002-2598-542X","position":2,"is_corresponding":false},{"id":459421,"name":"Monika Kizerwetter","orcid":"0000-0001-6084-7471","position":3,"is_corresponding":false},{"id":459422,"name":"Yanran He","orcid":"0000-0002-5814-5734","position":4,"is_corresponding":false},{"id":441176,"name":"Steven P. Wolf","orcid":"0009-0008-2723-695X","position":5,"is_corresponding":false},{"id":441173,"name":"Karin Schreiber","orcid":"0009-0000-1414-9365","position":6,"is_corresponding":false},{"id":296110,"name":"Henrik Clausen","orcid":"0000-0002-0915-5055","position":7,"is_corresponding":false},{"id":441180,"name":"Hans Schreiber","orcid":"0000-0002-1908-6233","position":8,"is_corresponding":false},{"id":297956,"name":"David M. Kranz","orcid":"0000-0002-4931-126X","position":9,"is_corresponding":false},{"id":459418,"name":"Preeti Sharma","orcid":"0000-0002-8210-9004","position":0,"is_corresponding":true}],"reference_count":58,"raw_metadata":null,"created_at":"2026-07-18T22:29:21.932251Z","pmid":"32541028","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":[]}