Electrophoresis Titration Model of a Moving Redox Boundary Chip for a Point-of-Care Test of an Enzyme-Linked Immunosorbent Assay.

Enzyme-linked immunosorbent assays (ELISAs) have been widely used in clinical examination, food safety, and environmental analyses. However, they still face a great challenge in designing a device for a point-of-care test (POCT) due to its bulk optical detector and complexity. Herein an electrophoresis titration (ET) model of a moving redox boundary (MRB) was proposed for constructing an ET-ELISA chip of a POCT just with sextuplet electrode pairs and laminated cells. The chip had an anodic well, middle well, and cathode well which were connected by microchannels. The ELISA process was conducted in the bottom of the middle well, where horseradish peroxidase (HRP) catalyzed 3,3',5,5'-tetra-methyl benzidine (TMB) as a blue TMB dimer with two positive charges. Under an electrical field of 29 V, the TMB dimer migrated into the titration channel and reacted with the ascorbic acid, creating an MRB. The MRB motion was a function of antigen content, indicating a visual distance-based assay. As a proof of concept, a C-reactive protein was chosen as a model antigen. The experiments systemically validated the ET-ELISA model and method. Particularly, the chip was smartphone-detected, traditional power supply free, and did not use sulfuric acid used in typical ELISA, making the ET-ELISA method extremely simple, portable, and safe. The ET-ELISA has great potential to visual and portable ELISA in clinical medicine, the environment, and food safety immunoassay.