Nano-LCMS in Routine Bioanalysis Application to Human NGF Biomarker Studies

Presented by: Gary A. Schultz, Ph.D., Senior Director, Biomarkers, Advion BioServices, Inc.

The number of monoclonal antibodies under development as therapeutic agents has risen significantly. Tracking the concentration profile of the total target biomarker during therapy provides an important contribution to understanding the pharmacokinetic/ pharmacodynamic relationship This presentation highlights the performance of an immunoaffinity nanoLC-MS/MS assay and the impact it has on clinical development programs of a therapeutic antibody, for which immunoassay development was unsuccessful. This assay quantifies total human β-nerve-growth factor (NGF) in serum by immunoprecipitation enrichment using a biotinylated human β-NGF affinity purified polyclonal antibody.

A 3-column liquid chromatography configuration is described with a cycle time of 10 min, combining a peptide immunoaffinity enrichment column with elution to nanoflow LC-MS/MS, providing a robust high sensitivity platform. Advanced assay validation was performed using a 7.0-450 pg/mL calibration range. Clinical phase I and II programs are supported with sample numbers ranging from a few hundred to more than seven thousand. The consistent implementation of this immunoaffinity LC-MS/MS assay to a high quality standard and with wide clinical impact is considered a tremendous technological achievement. The data now supports clinical dose selection, route of administration changes and systems pharmacology approaches.

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