HCP Identification and ELISA Coverage Methods in Novel Modalities

Contributing Authors: Johanna Barnhill, M.S., Stephen Stahlschmidt, Timothy Licknack, Ph.D., Alla Zilberman, Ph.D., and Jared Isaac, Ph.D. Research & Development Department, Chromatography and Mass Spectrometry Group Cygnus Technologies has performed antibody coverage analysis of over 600 unique samples to determine immunoreactive host cell proteins (HCPs) against antibody pools from dozens of different ELISA kits via Antibody Affinity Extraction (AAE). In USP 1132.1(1), AAE is referred to as “immunoaffinity chromatography” and as an accepted method for antibody coverage analysis. Applicable to all expression platforms (e,g,. CHO, E. coli, HeLa), diverse types of biotherapeutics (e.g., mAbs, viral vectors for gene therapies), and compatible with both two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) or liquid chromatography / mass spectrometry (LC-MS); AAE is a powerful tool for enriching HCPs. AAE data have been included in submissions to nearly all global regulatory bodies including the FDA, CFDA, MHRA, and other European and Asian regulatory authorities. Here, we briefly summarize over ten years of AAE experience.