Protein A chromatography: Challenges and progress in the purification of monoclonal antibodies

Abstract

Antibodies for therapeutic use are being continuously approved and their demand has been steadily growing. As known, the golden standard for monoclonal antibody (mAb) purification is Protein A affinity chromatography, a technology that has gained high interest because of its great performance and capabilities. The main concerns are the elevated resins costs and their limited lifetime compared to other resins (e.g. ion exchange chromatography). Great efforts have been carried out to improve purification conditions, such as resin characterization and designing alkali/acid stable resins with a longer lifetime. Modification of Protein A ligands and alternative formats such as monoliths membranes and microshperes have been tested to increase the purification performance. New technology has been proposed to improve the large‐scale separation; in addition, alternative ligands have been suggested to capture mAbs instead of Protein A ligand; however, most of the information is locked by pharmaceutical companies. This mini review summarizes and describes the advances, results, and impact on the Protein A chromatography purification processing.

Magnetic Precipitation: A New Platform for Protein Purification

“…As a standard capture for mAbs, protein A chromatography presents a high recovery yield and purity, usually above 95%. However, contrary to protein A chromatography where a harsh pH denaturing elution condition is used, [ 3 ] the combination of the precipitant condition with PEG, along with the elution condition with PBS makes this a biocompatible and mild capture process for mAb purification. [ 40,41 ] The elution fraction from protein A chromatography is many times directly used as the source for a viral inactivation step due to its low pH, but with no control regarding the pH‐lowering rate or mAb concentration that may lead to protein aggregation and precipitation.…”

“…[ 1 ] The purification of high added value biological drugs is mostly based on chromatographic methods. [ 2,3 ] Monoclonal antibodies (mAbs) and derived molecules are the most important players in the biopharmaceutical industry. [ 4 ] Albeit the advances in antibody engineering, antibody manufacturing still presents challenges.…”

On-column disulfide bond formation of monoclonal antibodies during Protein A chromatography eliminates low molecular weight species and rescues reduced antibodies

“…48 , 49 Additionally, it is critical to have an alternative resin in manufacturing with a demonstrated comparable performance to avoid any manufacturing disruption caused by a potential raw material supply shortage. 50 The possible mixture mode of binding of a different resin and mAb may cause a different molecular conformation and reoxidation behaviors (e.g., reaction kinetics and preferable pathways) that need further optimization of the redox condition. A comprehensive study is required to evaluate the alternative resin for the process performance and resultant mAb product quality.…”

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

“…Chromatography currently remains a basic tool in the manufacturing of biopharmaceuticals, more particularly downstream purification processes [261], despite an increasing interest in nonchromatographic techniques [262]. This prevalence is reflected in a number of recent reviews, covering for instance well-established AFC [263,264] and fast developing MMC [265,266], emerging materials -including AFC and MMC ligands and cellulose-based IEC media - [267], or else continuous chromatography [268].…”