Background
Bispecific antibodies (BsAb) belong to a novel antibody category with advantages over traditional mono-specific therapeutic antibodies. However, product variants are also commonly seen during the production of BsAb, which poses significant challenges to downstream processing. In this study, the adsorptive characteristics of a BsAb product and its variants were investigated for a set of depth filters during primary recovery of the cell culture fluid.
Conclusion
Harvest depth filters potentially can be utilized for retaining the BsAb variants, which depends on relative molecular properties of the product and its variants and adsorptive properties of the depth filters used.
Methods
The retention of the BsAb product and its variants on a set of Millistak+® D0HC and X0HC depth filters were first investigated, followed by studying the mechanism of their adsorption on the depth filters. The chemical and structural properties of depth filters along with the molecular properties of the product and its variants were studied subsequently.
Results
The X0HC filter was found to be able to retain a significant amount of low molecular weight (LMW) variants along with a low amount of main product retained. Different levels of retention, observed for these variants, were correlated to their different hydrophobic and charge characteristics in relation with the adsorptive properties of the depth filters used. Electrostatic, hydrophobic, and hydrogen bonding interactions were found to be the key forces to keep product variants retained on the depth filter where the higher hydrophobicity of the LMW variants may cause them to be preferentially retained.