Verifying SPARCL performance on the CLARIOstar equipped for reading at time of injection
SPARCL refers to an immunoassay that does not require washing steps. An analyte in a solution is detected by binding two antibodies: one is coupled to horse radish peroxidase (HRP), the other is coupled to the HRP substrate Acridan. In the presence of the analyte, binding of both antibodies brings the enzyme and substrate into proximity. The enzymatic reaction is started by the addition of H2O2, which immediately produces a light proportional to the amount of analyte. Thus, SPARCL assays require to be read at the same time as the injection of the trigger solution.
The CLARIOstar® microplate reader, which is equipped with the dedicated H3 injection needle for simultaneous injection and reading, fulfils this task. The analyte IgG was detected down to a concentration of 45 ng/ml in 96 wells and down to 15.6 ng/ml in 384 well format. BMG LABTECH's MARS Data Analysis Software facilitates subsequent calculation of analyte concentration.