Deep Profiling of Host Cell Proteins in Downstream Processing of Therapeutic Monoclonal Antibodies: Novel Approach to Isolate and Digest Host Cell Proteins

Host cell proteins (HCPs) are process-related impurities generated during the production of biopharmaceuticals. Due to their potential impact on product safety, quality and efficacy, regulatory authorities require removal of HCPs during processing down to trace amounts in final manufactured biopharmaceuticals.

This presentation will discuss a rapid and efficient workflow for the isolation, digestion and identification of HCPs. For the first time, Fc-receptor (FcγRIIIa) affinity chromatography was employed to isolate the HCP fraction from the mAb. Next, the HCPs were precipitated with acetone and digested using a newly developed “single-pot” method that improves digestion performance and prevents loss of problematic low-abundant HCPs. When compared to protein A chromatography, this method could reduce the analysis time by collecting a single fraction while increasing the number of identified HCPs. The strategy is applicable to a wide range of therapeutic mAbs and provides a reproducible, straightforward, easily implementable procedure that can be carried out in standard laboratories without the need for dedicated personnel or equipment.

Contributing Authors:

Jerome Jonveaux and Michael Jahn