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Competitive bead-based fluorescence assay to quantify antibody concentration

Christine Wosnitza, Sebastian Giehring, PAIA Biotech GmbH, Cologne, Germany; Franka Maurer BMG LABTECH GmbH, Ortenberg, Germany, 12/2015

Antibody quantification is the everyday work in cell line development labs. Thousands of cell clones lead to thousands of samples which need to be tested. The standard procedure for this determination is the time-consuming ELISA or Western Blot method. An assay allowing a higher throughput to save material and time is highly needed.

The company PAIA Biotech has developed a bead-based kit to determine the amount of antibody in samples in a high throughput format. Cell clones that have been grown in 384-well secrete its protein into the supernatant. Only a very low sample volume of 2-10 µl of the supernatant is needed to be used in the PAIA assays.

The innovative basis of the assay is a 384-well PAIA plate that has protrusions. This allows the separation of beads bound to either analyte or fluorescence marker without washing, blocking or regeneration steps. In this application note we show how the CLARIOstar microplate reader has been set up to measure the competitive human IgG Fc kit for the monoclonal antibodies Cetuximab, Rituximab and Panitumumab.