pH‐based cation exchange chromatography in the capture and elution of monoclonal antibodies

Abstract

Cation exchange chromatography separates adsorbed proteins by controlling the salt concentration or the mobile phase pH. This study examines the pH-based method for binding and elution of monoclonal antibodies (MAbs). Five different clones with isoelectric points from 6 to 9 were evaluated. We performed our studies using a new chromatography resin (Nuvia™ S), which has high binding capacity. A three-column process incorporating Nuvia S as a capture step was also demonstrated for the purification of MAb from tissue culture fluid. Chromatography performance of Nuvia S was demonstrated in a 50-cycle study.

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

“…IEX is the preferred technique for preparative and process scale separation for charge variants. While bench‐top analytical scale resolution has improved tremendously (Amand et al, ; Farnan & Moreno, ; Fekete et al, , ; Follman & Fahrner, ; Joshi, Kumar, & Rathore, ; Kang, Kutzko, Hayes, & Frey, ; Lingg et al, ; Ng & Snyder, ; Nordborg et al, ; Rea, Moreno, Lou, & Farnan, ; Santora, Kaymakcalan, Sakorafas, Krull, & Grant, ; Talebi et al, ; Talebi, Shellie, Hilder, Lacher, & Haddad, ; Tao, Carta, Ferreira, & Robbins, , ; Tao, Perez, Carta, Ferreira, & Robbins, ; Weitzhandler et al, ; Zhang, Patapoff, Farnan, & Zhang, ), industrial separation of charge variants remains challenging due to the scale and capacity required. For most industrial processes, the stationary phase, physical parameters of the columns, are already fixed.…”

Evaluation of a PEG/hydroxypropyl starch aqueous two‐phase system for the separation of monoclonal antibodies from cell culture supernatant

“…Abbreviations: ATPS, aqueous two-phase system; ATPE, aqueous two-phase extraction; CHO, Chinese hamster ovary; HPS, hydroxypropyl starch; IgG, immunoglobulin G; mAb, monoclonal antibody; VR, volume ratio regeneration, ligand leaching, and antibody aggregation at low pH elution conditions [6]. Recently, a variety of alternative techniques have been developed to improve the process efficiency and economics of antibody production, including mixed-mode adsorption [7], membrane adsorption [8], cationexchange chromatography [6], fluoroapatite chromatography [9], and aqueous two-phase extraction (ATPE).…”

“…Recently, a variety of alternative techniques have been developed to improve the process efficiency and economics of antibody production, including mixed-mode adsorption [7], membrane adsorption [8], cationexchange chromatography [6], fluoroapatite chromatography [9], and aqueous two-phase extraction (ATPE).…”

Cathepsin L Causes Proteolytic Cleavage of Chinese‐Hamster‐Ovary Cell Expressed Proteins During Processing and Storage: Identification, Characterization, and Mitigation

“…These include low‐binding capacity, high cost, lack of an effective cleaning procedure, or short lifetime. Nonaffinity capture methods (e.g., ion exchange and mixed mode) have been evaluated in several studies and separation of HCPs is often found to be challenging because of the nonspecific nature of these separation techniques . Therefore, an in‐depth understanding of the HCP chromatographic behavior would provide valuable insight for the rational design of purification processes …”

“…Nonaffinity capture methods (e.g., ion exchange and mixed mode) have been evaluated in several studies and separation of HCPs is often found to be challenging because of the nonspecific nature of these separation techniques. [14][15][16][17] Therefore, an in-depth understanding of the HCP chromatographic behavior would provide valuable insight for the rational design of purification processes. 18 Besides the immunogenicity concern, HCPs can also compromise product quality and shelf-life by resulting in degradation of protein or excipient, which leads to fragmentation, aggregation, and particle formation.…”