One-incubation one-hour multiplex ELISA enabled by aqueous two-phase systems. Issue 10 (6th April 2020)

• Record Type: Journal Article
• Authors: Tongdee, Mintra; Yamanishi, Cameron; Maeda, Midori; Kojima, Taisuke; Dishinger, John; Chantiwas, Rattikan; Takayama, Shuichi
• Is Part Of: Analyst. Volume 145:Issue 10(2020)
• Page Start: 3517
• Page End: 3527
• Publication Date: 2020-04-06
• Subjects: Chemistry, Analytic -- Periodicals; 543
• DOI: 10.1039/d0an00383b
• Languages: English
• ISSNs: 0003-2654

Abstract

Abstract : This work presents one-incubation one-hour multiplex ELISA enabled by aqueous two-phase systems for five-plex cytokine detection in human ThP-1 macrophages. 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.