Mixed Matrix PVDF Microfiltration Membranes with In-situ Synthesized Polyethyleneimine Particles as a Platform for Flow Through, High Capacity, Weak Base and Salt Tolerant Anion Exchange Membrane Adsorbers for Downstream Bioprocessing

Abstract

The rapid growth of the monoclonal antibody (mAb) therapeutic market has led to a need for improved downstream bioprocessing, including mAb capture and release from bioreactor harvests followed by product purification and polishing. Membrane chromatography (MC) is poised to displace resin-based column chromatography in mAb product polishing. To remove negatively charged product impurities (e.g., host cell proteins, DNA, viruses, and endotoxins), anion exchange (AEX) membrane adsorbers offer higher process rates and eliminate the risks of cross contamination as they can be deployed as single use separation devices, which are increasingly being utilized in downstream bioprocessing to increase manufacturing speed and decrease production cost. Here, we describe a one-pot and single step phase inversion casting process for the preparation of a family of mixed matrix polyvinylidene fluoride (PVDF) microfiltration (MF) membranes with in-situ synthesized polyethyleneimine (PEI) microparticles that can serve as flow through, high capacity, and salt tolerant weak base AEX membrane adsorbers. These new membranes have a high content of weak base (WB) groups (primary and secondary amines) by virtue of the utilization of bis(2-chloroethyl)amine hydrochloride (BCAH) as crosslinker in the in-situ synthesis of PEI microparticles in the dope dispersions prior to membrane casting. By varying the concentration of PEI and BCAH in the casting dispersions, we successfully utilized nonsolvent induced phase separation (NIPS) with isopropyl alcohol as the nonsolvent in the membrane coagulation bath to prepare a mixed matrix PVDF-PEI MF membrane with (i) a relatively uniform and open microstructure in which the PEI particles completely cover the PVDF spherulites present at the membrane surface and cross section, (ii) high water flux (>1000 liters/m2/hr. at 2 bar) and (iii) a high loading of BCAH crosslinked PEI microparticles (51.0 wt.%) containing weak base primary, secondary, and tertiary amine groups. The protein binding measurements show that this new mixed matrix PVDF-PEI MF membrane can serve as a flow through, high capacity, and salt tolerant WB AEX membrane adsorber with a bovine serum albumin (BSA) dynamic binding capacity of ∼60 mg BSA per mL of membrane in a 50 mM TRIS buffer containing 100 mM of NaCl (15 mS/cm) at 10% breakthrough and flow rate of 10 MV/min. The overall results of this study indicate that our mixed matrix PVDF-PEI MF membranes with in-situ synthesized and BCAH crosslinked PEI microparticles have a promising potential to serve as a platform for the design, preparation, and scale up of a new generation of flow through, high capacity, salt tolerant, WB AEX membrane adsorber materials for downstream bioprocessing.