Abstract

Magnetic bead immunocapture-LC-MS has been widely used for bioanalysis of biotherapeutic proteins. However, magnetic beads are difficult to be fully automated and more costly than ELISA plates. Develop an ELISA-LC-MS hybrid assay as an alternate platform. Among seven ELISA plates tested, Pierce streptavidin plates, which did not require time-consuming capture antibody precoating steps, provided the best sensitivity and assay dynamic range (5-2500ng/ml or 10-5000ng/ml), similar to magnetic bead immunocapture-LC-MS assay and better than an ELISA (50-500ng/ml). The entire procedures could be fully automated using a liquid handling system. This study demonstrates that ELISA-LC-MS hybrid approach using streptavidin plates represents a promising platform for bioanalysis of biotherapeutics.

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