{"doi":"10.1021/acsomega.2c03237","title":"A Rapid Method for Direct Quantification of Antibody Binding-Site Concentration in Serum","abstract":"The quantitation of the available antibody binding-site concentration of polyclonal antibodies in serum is critical in defining the efficacy of vaccines against substances of abuse. We have conceptualized an equilibrium dialysis (ED)-based approach coupled with fluorimetry (ED-fluorimetry) to measure the antibody binding-site concentration to the ligand in an aqueous environment. The measured binding-site concentrations in monoclonal antibody (mAb) and sera samples from TT-6-AmHap-immunized rats by ED-fluorimetry are in agreement with those determined by a more established equilibrium dialysis coupled with ultraperformance liquid chromatography tandem mass spectrometry (ED-UPLC-MS/MS). Importantly, we have shown that the measured antibody binding-site concentrations to the ligand by ED-fluorimetry were not influenced by the sample serum matrix; thus, this method is valid for determining the binding-site concentration of polyclonal antibodies in sera samples. Further, we have demonstrated that under appropriate analytical conditions, this method resolved the total binding-site concentrations on a nanomolar scale with good accuracy and repeatability within the microliter sample volumes. This simple, rapid, and sample preparation-free approach has the potential to reliably perform quantitative antibody binding-site screening in serum and other more complex biological fluids.","journal":"ACS Omega","year":2022,"id":290236,"datarank":0.24141568686511508,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"self_citation_contribution":0.24141568686511508,"citation_network_contribution":0.0,"self_endowment_contribution":0.24141568686511508,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9597,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":625063,"name":"Connor Whalen","orcid":null,"position":1,"is_corresponding":false},{"id":518848,"name":"Oscar B. Torres","orcid":"0000-0002-5352-4647","position":2,"is_corresponding":false},{"id":972272,"name":"Alexander James Duval","orcid":"0000-0001-7540-4008","position":3,"is_corresponding":false},{"id":415888,"name":"Agnieszka Sulima","orcid":"0000-0002-8849-9736","position":4,"is_corresponding":false},{"id":958154,"name":"Joshua F. G. Antoline","orcid":null,"position":5,"is_corresponding":false},{"id":625061,"name":"Therese Oertel","orcid":null,"position":6,"is_corresponding":false},{"id":336553,"name":"Rodell C. Barrientos","orcid":"0000-0002-6006-5047","position":7,"is_corresponding":false},{"id":439445,"name":"Arthur E. Jacobson","orcid":"0000-0002-0011-8354","position":8,"is_corresponding":false},{"id":301852,"name":"Kenner C. Rice","orcid":"0000-0002-1147-9147","position":9,"is_corresponding":false},{"id":332553,"name":"Gary R. Matyas","orcid":"0000-0002-2074-2373","position":10,"is_corresponding":false},{"id":972271,"name":"Erwin G. Abucayon","orcid":"0000-0003-0872-0857","position":0,"is_corresponding":true}],"reference_count":42,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T00:30:26.667578Z","pmid":"35936462","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":[]}