Background: 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. Aim: Develop an ELISA–LC–MS hybrid assay as an alternate platform. Results: 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 (5–2500 ng/ml or 10–5000 ng/ml), similar to magnetic bead immunocapture–LC–MS assay and better than an ELISA (50–500 ng/ml). The entire procedures could be fully automated using a liquid handling system. Conclusion: This study demonstrates that ELISA–LC–MS hybrid approach using streptavidin plates represents a promising platform for bioanalysis of biotherapeutics.
Keywords:
- ELISA
- hybrid assay
- immunocapture
- LC–MS
- magnetic beads
- monkey
- plasma
- streptavidin plate