Protein A chromatography remains a critical step in the purification of mAb and related products.  Its ability to remove >98 % of impurities alleviates the burden on subsequent steps and facilitates platform processing.  We have evaluated the ability of four commercially available Protein A chromatography matrices to remove host cell proteins (HCPs), based on a scale-down process for an IgG4 produced in CHO cells.  SELDI-TOF mass spectrometry was used to generate a contaminant profile for each resin and demonstrated that a number of components are shared, but further peaks are associated with porous glass.  To further examine the behaviour of these contaminants a null cell line approach was used to prepare enriched samples of HCPs for 2D-gel electrophoresis. Some contaminants are associated with the resin backbone while others with the product.  This characterisation may provide a rationale for targeted process development or cell line engineering strategies to remove challenging contaminants.

Characteristics of host cell protein removal during Protein A chromatography

243rd National Meeting (2012)

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