SimpleStep ELISA™: Innovative immunoassays in less than 90 minutes (TECH2P.918)

Abstract

Abstract Here we introduce the novel SimpleStep ELISA™ platform which has improved features over the traditional sandwich assay. The SimpleStep ELISA format is a novel, streamlined protocol that can be completed in less than 90 minutes, requires no additional training and uses standard laboratory equipment. In one step, the complete sandwich complex forms in solution and is anchored to the plate through an immunoaffinity tag. Only a single incubation and wash step is required, compared to multiple incubation and wash steps with standard ELISAs, that can take up to five hours. Fewer steps mean less time is needed and there are fewer opportunities for errors with no loss on sensitivity or increased background. While simple in concept, ELISA performance can be affected by many factors, including the sample type, as many antibody pairs and immunoassays that work well on purified proteins fail to perform acceptably when a complex biological sample, such as plasma or serum, is used. The foundation of SimpleStep ELISA performance is an extensive screening program to identify antibody pairs that perform well in a range of biological samples, and the inclusion of specially-designed buffers to ensure accuracy and minimize the effect of non-specific binding, for example by human serum antibodies reactive against mouse antibodies. Supporting data for each SimpleStep ELISA includes sensitivity, range, precision, linearity, recovery and cross-reactivity.