Membrane Adsorber for the Fast Purification of a Monoclonal Antibody Using Protein A Chromatography

Abstract

Monoclonal antibodies are conquering the biopharmaceutical market because they can be used to treat a variety of diseases. Therefore, it is very important to establish robust and optimized processes for their production. In this article, the first step of chromatography (Protein A chromatography) in monoclonal antibody purification was optimized with a focus on the critical elution step. Therefore, different buffers (citrate, glycine, acetate) were tested for chromatographic performance and product quality. Membrane chromatography was evaluated because it promises high throughputs and short cycle times. The membrane adsorber Sartobind® Protein A 2 mL was used to accelerate the purification procedure and was further used to perform a continuous chromatographic run with a four-membrane adsorber-periodic counter-current chromatography (4MA-PCCC) system. It was found that citrate buffer at pH 3.5 and 0.15 M NaCl enabled the highest recovery of >95% and lowest total aggregate content of 0.26%. In the continuous process, the capacity utilization of the membrane adsorber was increased by 20%.

Single-step rapid chromatographic purification and characterization of clinical stage oncolytic VSV-GP

“…Cation exchange chromatography is a purification platform used frequently for purification of not only proteins and antibodies (Brämer et al, 2019;Boi et al, 2020), but can also be efficiently utilized for the purification of complex viruses, as demonstrated in this study and previously by other authors (Eckhardt et al, 2022;Mueller et al, 2022). For example, Eckhardt et al (Eckhardt et al, 2022) have used resin based cation exchange chromatography for the purification of measles virus.…”

Matrices and Affinity Ligands for Antibody Purification and Corresponding Applications in Radiotherapy

“…As an alternative to column chromatography, membrane chromatography that combines membrane filtration and liquid chromatography together has captured growing attention for antibody purification. Membranes offer the advantages of simplicity, ease of handling, a larger surface area, an improved mass-transfer efficiency, and an easier scale-up [ 21 , 22 ]. Affinity ligands, such as protein A/G [ 21 ], mimetic™ A2P [ 23 ], ligand 22/8 [ 24 ], and tryptamine [ 25 ], have been immobilized onto membranes for antibody purification from complex mixtures.…”

“…Membranes offer the advantages of simplicity, ease of handling, a larger surface area, an improved mass-transfer efficiency, and an easier scale-up [ 21 , 22 ]. Affinity ligands, such as protein A/G [ 21 ], mimetic™ A2P [ 23 ], ligand 22/8 [ 24 ], and tryptamine [ 25 ], have been immobilized onto membranes for antibody purification from complex mixtures. For work regarding a detailed review of affinity membranes, please refer to the following article [ 22 ].…”

Developments and opportunities in continuous biopharmaceutical manufacturing

“…A continuous four-membrane adsorber periodic countercurrent chromatography system with increased throughput compared to Protein A column chromatography has been reported. 73 In addition, there have been considerable increases in capacity in newer generations of adsorptive membranes.…”