Enhanced Tangential Flow Filtration of Precipitated Proteins Using Screened Membrane Cassettes

Recent advances in cell culture have led to significant increases in monoclonal antibody (mAb) titers, opening a new window of opportunity for developing a fully continuous downstream purification process based on the selective precipitation of the mAb from harvested cell culture fluid, with the precipitate dewatered and washed using single-pass tangential flow filtration (SPTFF) with microfiltration membranes.

Experiments were performed with precipitates of human serum immunoglobulin G formed using ZnCl₂ and polyethylene glycol, both with and without added disodium malonate. SPTFF was conducted in both hollow fiber and screened cassette modules, with the critical flux identified using flux-stepping experiments.

Critical fluxes as high as 250 L/m²/h were obtained in the screened cassette, significantly higher than what was possible in hollow fiber modules. A two-stage system was designed that provided up to 85% conversion in a single pass. This system could be operated continuously for 24 h with 80% conversion at a filtrate flux of 144 L/m²/h without any significant fouling.

The results demonstrate the potential of using screened membrane cassettes for the continuous/intensified processing of precipitated proteins like monoclonal antibodies.