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Assay Development for High Content Quantification of Sod1 Mutant Protein Aggregate Formation in Living Cells
Published on: October 4, 2017
Development of an ELISA-LC-MS hybrid assay for quantification of biotherapeutics
Linzhi Chen1, Siyu Liu1,2, Shirin Pagels1
1Drug Metabolism & Pharmacokinetics, Boehringer Ingelheim Pharmaceuticals, Ridgefield, CT 06877.
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.
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