High-throughput, semi-homogeneous enzyme-linked immunosorbent assays in 384-well plates, ready to analyze in 90 minutes

Abstract: After more than 40 years, the enzyme-linked immunosorbent assay (ELISA) is still considered to be one of the most reliable assays, and often considered a gold standard, for the detection and quantification of proteins and other analytes in various biological samples. Traditional ELISA methods require several wash steps and relatively long turn-around times (3-5 hours) as well as high sample volumes, making them more challenging to adapt to high-throughput processes. Abcam’s SimpleStep ELISA technology streamlines the process to a semi-homogeneous format that results in a simple 90-minute, single-wash protocol. For higher throughput demands, SimpleStep ELISA can be miniaturized to a 384-well format allowing up to 4 times more samples to be processed in the same amount of time.  Here we describe how SimpleStep ELISa in 384-well plates can be used to quantify human IgG and human Interferon gamma (IFN gamma), two important targets in pharmacokinetic studies and immunotherapeutic drug safety testing. A microplate reader equipped with a spectrometer for absorbance measurements produced high precision results in with read times of less than one second per well. This, combined with the cost-savings provided by miniaturization to 50 μl, provide an assay suitable for high-throughput.