Iminodiacetic Acid (IDA) Cation-Exchange Nonwoven Membranes for Efficient Capture of Antibodies and Antibody Fragments

Abstract

There is strong need to reduce the manufacturing costs and increase the downstream purification efficiency of high-value therapeutic monoclonal antibodies (mAbs). This paper explores the performance of a weak cation-exchange membrane based on the coupling of IDA to poly(butylene terephthalate) (PBT) nonwoven fabrics. Uniform and conformal layers of poly(glycidyl methacrylate) (GMA) were first grafted to the surface of the nonwovens. Then IDA was coupled to the polyGMA layers under optimized conditions, resulting in membranes with very high permeability and binding capacity. This resulted in IgG dynamic binding capacities at very short residence times (0.1–2.0 min) that are much higher than those achieved by the best cation-exchange resins. Similar results were obtained in the purification of a single-chain (scFv) antibody fragment. As is customary with membrane systems, the dynamic binding capacities did not change significantly over a wide range of residence times. Finally, the excellent separation efficiency and potential reusability of the membrane were confirmed by five consecutive cycles of mAb capture from its cell culture harvest. The present work provides significant evidence that this weak cation-exchange nonwoven fabric platform might be a suitable alternative to packed resin chromatography for low-cost, higher productivity manufacturing of therapeutic mAbs and antibody fragments.

Purification of Adeno-Associated Virus (AAV) Serotype 2 from Spodoptera frugiperda (Sf9) Lysate by Chromatographic Nonwoven Membranes

“…Sartobind Q pico 0.08 mL was purchased from Sartorius (Goettingen, Germany). CEX-IDA membrane was fabricated following the method developed in [ 38 ]. The clarified Sf9 cell lysate and a solution of pure AAV2 carrying a green fluorescent protein (GFP) transgene were purchased from Virovek (Hayward, CA, USA).…”

“…A scanning electron microscope (SEM, Hitachi S-3200 N, Hitachi High-Tech, Schaumburg, IL, USA) was used to image the membrane structure after preparation. The CEX-IDA membrane was prepared following the process established in published work [ 38 ].…”

“…Our team has developed a class of nonwoven membrane adsorbers for the chromatographic purification of biologics, comprising polybutylene terephthalate (PBT) nonwoven fabrics grafted with polyglycidyl methacrylate (GMA) layers that can be functionalized with a variety of ligands [ 36 , 37 , 38 ]. These membranes exhibit excellent flow permeability, high binding capacity and selectivity towards proteins, and can be operated in bind-and-elute mode for product capture or flow-through mode for product polishing.…”

“…For this purpose, nonwoven membranes UV grafted with glycidyl methacrylate (GMA) were functionalized with (i) triethylamine (TEA) to form a quaternary ammonium anion-exchange membrane (AEX-TEA membrane), and (ii) iminodiacetic acid (IDA), a carboxylic acid ligand, to form a cation-exchange membrane (CEX-IDA membrane). While the AEX-TEA membrane is a new anion exchanger, the CEX-IDA membrane was previously developed for the purification of monoclonal antibodies [ 38 ], although it was never applied in the purification of viral vectors.…”

Designing polymeric membranes with coordination chemistry for high-precision ion separations

“…Here, we demonstrate how cation permeability and selectivity are affected by coordinative interactions with a polymer membrane. Our analysis focuses on transport of similar metal cations through polyelectrolyte multilayer membranes with iminodiacetate (IDA) functional groups, owing to their strong coordinative interactions ( 21 , 25 , 26 ). Results demonstrate that metal ions with higher binding energy to IDA groups more selectively pass through the membrane in multisalt solutions than in single-salt solutions, whereas the permeability of weaker binding species is markedly reduced because they are less likely to bind with IDA moieties.…”