Automating Sample Preparation Workflows for Hybrid LC-MS/MS Bioanalysis of Protein Therapeutics: Quantification of Etanercept Using Affinity Purification and Digestion

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Paula Orens and Mary Lame
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Application Notes
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This application note aims to describe the process of developing a successful, automated capture and digestion method for hybrid LC-MS/MS quantification of the fusion protein, etanercept.

Prior to quantification, the comparability of the automated and manual preparations was demonstrated in a step wise approach, assessing the critical steps of the protocol individually. This limited the variables during automation optimization and ultimately decreased the time spent on script development. This approach to developing an automated capture and digestion sample preparation facilitated accurate and reproducible quantification of etanercept from rat plasma down to 1 ng/mL.

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