P-115-R: Global HCP Profiling and Quantifications by inSPECt™ LC-MS Assay

Liquid Chromatography-Mass Spectrometry (LC-MS) is a powerful tool for identifying host cell proteins (HCPs), but most high-throughput workflows often yield inaccurate results. A new LC-MS workflow was developed and qualified to identify and quantify up to hundreds of HCPs in downstream biopharmaceutical samples, called the inSPECt MS Assay. InSPECt utilizes native digestion for sample preparation and Data-Independent Acquisition using the state-of-the-art Astral Zoom Mass Spectrometer (Thermo Fisher Scientific). To evaluate the analytical limits of this method, calibration curves with spike-in protein standards were generated; subsequently, a unique blend of problematic HCPs were spiked into a representative Drug Substance (DS) sample to evaluate method performance. Our results indicate that HCP inSPECt is an accurate and precise assay for identification and quantification of HCPs in process samples. Contributing Authors: Timothy Licknack, Stephen Stahlschmidt, Johanna Barnhill, Brinkley Thornton, Jared Isaac Cygnus Technologies, Leland, United States