{"doi":"10.1002/bit.28954","title":"Engineering Affibody Binders to Death Receptor 5 and Tumor Necrosis Factor Receptor 1 With Improved Stability","abstract":"ABSTRACT Protein developability is an important, yet often overlooked, aspect of protein discovery campaigns that is a key driver of utility. Recent advances have improved developability screening capacity, making it an increasingly viable option in early‐stage discovery. Here, we engineered one component of developability, stability, of two affibody proteins—one that targets death receptor 5 and another that targets tumor necrosis factor receptor 1—previously evolved to bind receptor and non‐competitively inhibit signaling via conformational modulation. Starting from an error‐prone PCR library of each affibody, variants were screened via yeast surface display binder selections, including depletion of non‐specific binders, followed by developability assessment using the on‐yeast protease and yeast display level assays. Multiplex deep sequencing identified variants for further evaluation. Purified variants exhibited elevated stability—8°C to 14°C increase in T m,app —with maintained 1–2 nM affinity for the TNFR1 affibody and 30‐fold improvement in the DR5 affibody affinity to 0.8 nM.","journal":"Biotechnology and Bioengineering","year":2025,"id":562820,"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":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9458,"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":483417,"name":"Jonathan N. Sachs","orcid":"0000-0003-1403-5960","position":1,"is_corresponding":false},{"id":408882,"name":"Benjamin J. Hackel","orcid":"0000-0003-3561-9463","position":2,"is_corresponding":false},{"id":1012122,"name":"Gregory H. Nielsen","orcid":null,"position":0,"is_corresponding":true}],"reference_count":41,"raw_metadata":null,"created_at":"2026-07-19T02:56:09.638214Z","pmid":"40045532","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":[]}