{"doi":"10.1016/j.mtbio.2025.102698","title":"Conjugation strategy shapes antitumor efficacy and enables dose-sparing in non-antibody protein nanoconjugates","abstract":"Precision targeting is a hot topic in cancer nanomedicine, as conventional chemotherapies cause systemic toxicities, creating an urgent need for more selective treatments. Although antibody-drug conjugates (ADCs) are the current gold standard in targeted therapy, their clinical performance remains limited. As an alternative, we previously developed a multivalent protein nanocarrier (T22-GFP-H6) displaying the CXCR4-targeting peptide T22, which offers super-selective tumor accumulation driven by CXCR4 overexpression. This innovative nanovehicle showed favorable biodistribution for targeted delivery of antitumor drugs, including monomethyl auristatin E (MMAE), in a first-generation stochastic nanoconjugate format. However, unlike ADCs, where conjugation strategy is known to influence pharmacokinetics and efficacy, these parameters remain largely unexplored in non-antibody multivalent nanocarriers. Here, we evaluated the impact of precise payload accommodation using two site-specific strategies that attach a single MMAE molecule at distinct structural sites, and we compared them with first-generation nanoconjugates. The conjugation strategy substantially affected the biodistribution and antitumor efficacy, with a solvent-exposed cysteine-conjugation distal to the targeting ligand proving most effective. At equimolar nanocarrier dosing, this construct achieved tumor control similar to the stochastic conjugate in a disseminated hematologic malignancy despite an approximately 4-fold lower MMAE load (drug-to-protein ratio, DPR = 1 vs DPR ≈ 4). Moreover, at equimolar MMAE dosing, it clearly outperformed both the stochastic conjugate and the alternative site-directed design. These findings align with trends in advanced ADCs and provide practical design rules for rational, site-specific conjugation in next-generation protein-based nanomedicines aimed at enabling dose-sparing in oncology.","journal":"Materials Today Bio","year":2025,"id":585790,"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":0.9578,"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":933144,"name":"Annabel García‐León","orcid":"0000-0003-3007-9306","position":1,"is_corresponding":false},{"id":1500003,"name":"Lourdes A. Arena","orcid":null,"position":2,"is_corresponding":false},{"id":921494,"name":"Julián I. Mendoza","orcid":"0000-0002-1975-3606","position":3,"is_corresponding":false},{"id":371246,"name":"Anna Aviñó","orcid":"0000-0003-3047-738X","position":4,"is_corresponding":false},{"id":1499639,"name":"Carme Fàbrega","orcid":"0000-0003-3816-7634","position":5,"is_corresponding":false},{"id":371252,"name":"Ramón Eritja","orcid":"0000-0001-5383-9334","position":6,"is_corresponding":false},{"id":1141318,"name":"David Páez","orcid":"0000-0002-5596-6588","position":7,"is_corresponding":false},{"id":403014,"name":"Lorena Alba‐Castellón","orcid":"0000-0003-3449-7820","position":8,"is_corresponding":false},{"id":371253,"name":"Esther Vázquez","orcid":"0000-0003-1052-0424","position":9,"is_corresponding":false},{"id":371254,"name":"Antonio Villaverde","orcid":"0000-0002-2615-4521","position":10,"is_corresponding":false},{"id":371251,"name":"Ramón Mangues","orcid":"0000-0003-2661-9525","position":11,"is_corresponding":false},{"id":371250,"name":"Isolda Casanova","orcid":"0000-0002-1196-4724","position":12,"is_corresponding":false},{"id":371255,"name":"Ugutz Unzueta","orcid":"0000-0001-5119-2266","position":13,"is_corresponding":false},{"id":933145,"name":"Ariana Rueda","orcid":"0000-0001-7588-9279","position":0,"is_corresponding":true}],"reference_count":75,"raw_metadata":null,"created_at":"2026-07-19T02:59:24.273134Z","pmid":"41560806","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":[]}