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%.
Highlights
Monoclonal antibodies “deliver considerable medical benefits” [1]
No significant difference in chromatography performance was observed, standard PBS with pH 7.4 was chosen for monoclonal antibody binding
The elution in Protein A chromatography is much more critical as a low pH value may lead to aggregation and denaturation of the antibody [32,33]
Summary
Monoclonal antibodies (mAb) “deliver considerable medical benefits” [1]. B-cells as an immune response to the intrusion of antigens. They belong to the immunoglobulin family and are divided into five immunoglobulin classes (IgG, IgA, IgM, IgD and IgE), which are subdivided into subgroups (e.g., IgG1, IgG2, IgG3, IgG4). The light chains (LC) and heavy chains (HC) have a variable region (V-region) at one end that serves as a binding site for the antigen [4,5,6]. Monoclonal antibodies bind to a defined epitope of an antigen and trigger the immune defense system; this has attracted interest in their medicinal applications [4].
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