{"doi":"10.1039/d0an00383b","title":"One-incubation one-hour multiplex ELISA enabled by aqueous two-phase systems","abstract":"This work describes a convenient one-hour enzyme-linked immunosorbent assay (ELISA) formulated with conventional antibodies and horseradish peroxidase (HRP) reagents. The method utilizes aqueous two-phase system (ATPS) droplet formation based on poly(ethylene glycol) (PEG)-containing sample solution-triggered rehydration of dehydrated dextran (DEX) spots that contain all antibody reagents. Key advances in this paper include development of a formulation that allows a quick 1-hour overall incubation time and a procedure where inclusion of the HRP reagent in the PEG solution reduces the number of washing and incubation steps required to perform this assay. As an assay application, a 5-plex cytokine test compares cytokine secretion of differentially-treated human ThP-1 macrophages. Given the use of only readily available reagents and a common Western blot imaging system for the readout, this method is envisioned to be broadly applicable to a variety of multiplex immunoassays. To facilitate broader use, companion image processing software as an ImageJ plugin is also described and provided.","journal":"The Analyst","year":2020,"id":77252,"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":19,"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":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":402430,"name":"Cameron Yamanishi","orcid":"0000-0001-9190-3448","position":1,"is_corresponding":false},{"id":403510,"name":"Midori Maeda","orcid":null,"position":2,"is_corresponding":false},{"id":402431,"name":"Taisuke Kojima","orcid":"0000-0001-7956-5378","position":3,"is_corresponding":false},{"id":403511,"name":"John F. Dishinger","orcid":null,"position":4,"is_corresponding":false},{"id":402432,"name":"Rattikan Chantiwas","orcid":"0000-0002-3788-0908","position":5,"is_corresponding":false},{"id":282621,"name":"Shuichi Takayama","orcid":"0000-0002-4385-9080","position":6,"is_corresponding":false},{"id":402429,"name":"Mintra Tongdee","orcid":"0000-0003-3749-8650","position":0,"is_corresponding":true}],"reference_count":39,"raw_metadata":null,"created_at":"2026-07-18T21:48:50.085103Z","pmid":"32248215","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":[]}