Protein A chromatography is a critical and 'gold-standard' step in the purification of monoclonal antibody (mAb) products. However, the purification of target proteins can be challenging due to the co-purification of a complex group of process related host cell proteins (HCPs). Here, we have evaluated two commercially available protein A chromatography resins, Eshmuno A and ProSep Ultra Plus (PUP) with two different mAbs A and B respectively, in terms of their ability and process conditions to remove HCP. Different intermediate buffers and elution conditions were used to screen the best condition according to the HCP removal during protein A chromatography. We alter intermediate wash pH, buffer salt concentration and different type of additive, including detergent and solvent, and also the elution pH to generate the impurity profile fingerprint for each resin. The results showed the HCP removal capacity raise about 47 % and 111 % for Eshmuno A and PUP respectively result from the optimization process. And the consequence support that increasing elution pH was able to minimize HCP level while maximize target protein yield. It was also found that multi-component intermediate wash buffer was more effective in reducing HCP than the single component buffer which imply the HCP related interaction may not only be hydrophilic type but also hydrophobic or even amphoteric. The methods shown in this study allow for a critical remove of HCP during protein A chromatography which alleviates the burden on subsequent process steps and facilitates the implementation of platform processes. On the whole it provides a better solution for process engineers to optimize rational approaches for the removal of prominent HCPs for monoclonal antibody product.

Systemic approach to remove host cell proteins during monoclonal antibody protein A chromatography

255th National Meeting (2018)

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