{"doi":"10.1039/d5lc00745c","title":"Online affinity micro free-flow electrophoresis for the continuous monitoring of insulin <i>via</i> a competitive immunoassay","abstract":"An online, micro free-flow electrophoresis (μFFE) assay was developed for the real-time measurement of insulin in a continuous, competitive immunoassay. Fluorescently labeled insulin and anti-insulin monoclonal antibody were mixed with the sample stream online and flowed through an incubation capillary into the μFFE device. Under an electric field, the bound antibody : insulin-FITC complex was separated from free insulin-FITC. When unlabeled insulin was introduced, labeled and unlabeled insulin competed for binding sites on the antibody, and the separated peaks of the bound complex and free insulin-FITC responded in real-time. Temporal resolution of 30 seconds was achieved following injections of unlabeled insulin, with a limit of detection of 63 nM and a linear response over the tested range of 50 nM to 1 μM insulin. The development of this assay supports the eventual integration of an organ-on-a-chip system capable of measuring direct cellular efflux in tandem with the affinity μFFE detection system. To approximate such an online cellular response, glucose-stimulated insulin secretion was collected offline from human islets and injected into the affinity μFFE instrument. A measurable change in insulin secretion was observed between samples exposed to low and high glucose, demonstrating sufficient limit of detection, linearity, the compatibility of the instrument with a complex biological matrix.","journal":"Lab on a Chip","year":2025,"id":584475,"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":0.9605,"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":383332,"name":"Seokwon Jo","orcid":"0000-0002-8989-4893","position":1,"is_corresponding":false},{"id":232435,"name":"Emilyn U. Alejandro","orcid":"0000-0002-7941-8439","position":2,"is_corresponding":false},{"id":1129467,"name":"Michael T. Bowser","orcid":"0000-0002-1570-2414","position":3,"is_corresponding":false},{"id":275279,"name":"Gretchen S. Burke","orcid":"0009-0004-9474-7259","position":0,"is_corresponding":true}],"reference_count":56,"raw_metadata":null,"created_at":"2026-07-19T02:59:11.978098Z","pmid":"41576240","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":[]}