Pressure: a novel tool for enzyme-linked immunosorbent assay procedure

Abstract

The enzyme-linked immunosorbent assay (ELISA) technique is the backbone of the diagnostic assay for detection of infectious and allergic diseases. Here, we demonstrate a unique ELISA method for the detection of an antigen or antibody, in which ELISA steps are carried out under pressure instead of conventional thermal incubation. Pressure-mediated ELISA (PELISA), carried out in 1 h shows more than a 2-fold increase in absorbance value than the control experiment carried out at the same time and temperature without applying pressure. Estimation of total IgE by the 1-h PELISA method gives similar absorbance value (1.081±0.031, 823.12 IU) to that obtained by 3-h heat-mediated ELISA on an activated surface (HELISA) (1.165±0.037, 810.96 IU). Since PELISA is sensitive, specific, and reproducible (intra- and interassay CVs were 6.47% and 9.65%, respectively), it could be an excellent alternative to HELISA or conventional ELISA procedures.

Keywords

• enzyme-linked immunosorbent assay
• pressure-mediated ELISA
• 1-flouro-2-nitro-4-azidobenzene
• activated surface
• covalent immobilization