{"doi":"10.1016/j.jpha.2024.100966","title":"Mapping conformational changes on bispecific antigen-binding biotherapeutic by covalent labeling and mass spectrometry","abstract":"Biotherapeutic's higher order structure (HOS) is a critical determinant of its functional properties and conformational relevance. Here, we evaluated two covalent labeling methods: diethylpyrocarbonate (DEPC)-labeling and fast photooxidation of proteins (FPOP), in conjunction with mass spectrometry (MS), to investigate structural modifications for the new class of immuno-oncological therapy known as bispecific antigen-binding biotherapeutics (BABB). The evaluated techniques unveiled subtle structural changes occurring at the amino acid residue level within the antigen-binding domain under both native and thermal stress conditions, which cannot be detected by conventional biophysical techniques, e.g., near-ultraviolet circular dichroism (NUV-CD). The determined variations in labeling uptake under native and stress conditions, corroborated by binding assays, shed light on the binding effect, and highlighted the potential of covalent-labeling methods to effectively monitor conformational changes that ultimately influence the product quality. Our study provides a foundation for implementing the developed techniques in elucidating the inherent structural characteristics of novel therapeutics and their conformational stability.","journal":"Journal of Pharmaceutical Analysis","year":2024,"id":479698,"datarank":0.12298384290809544,"base_score":0.6931471805599453,"endowment":0.6931471805599453,"self_citation_contribution":0.10397207708399181,"citation_network_contribution":0.019011765824103632,"self_endowment_contribution":0.10397207708399181,"citer_contribution":0.019011765824103632,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":1,"citers_with_citation_signal":1,"citers_with_endowment":1,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9562,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1318766,"name":"Dipanwita Batabyal","orcid":"0009-0007-9791-7067","position":1,"is_corresponding":false},{"id":1319111,"name":"Dayong Qiu","orcid":null,"position":2,"is_corresponding":false},{"id":1096216,"name":"Weidong Cui","orcid":"0000-0002-2871-9485","position":3,"is_corresponding":false},{"id":1096540,"name":"John Harrahy","orcid":null,"position":4,"is_corresponding":false},{"id":226931,"name":"Alexander R. Ivanov","orcid":"0000-0002-4691-8488","position":5,"is_corresponding":false},{"id":1096215,"name":"Arnik Shah","orcid":"0000-0003-3455-0177","position":0,"is_corresponding":true}],"reference_count":88,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:06:54.284166Z","pmid":"39263356","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":[]}