Using hydrogen peroxide to prevent antibody disulfide bond reduction during manufacturing process
| Autor: | Joon Chong Yee, Nesredin Mussa, Zhijun Tan, Nripen Singh, Anthony J. Cura, Cheng Du, Zheng Jian Li, Ameya Borwankar, Michael C. Borys, Sanchayita Ghose, Yunping Huang, Richard Ludwig |
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| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Immunology Mixing (process engineering) chemistry.chemical_element CHO Cells 01 natural sciences Oxygen Chromatography Affinity 03 medical and health sciences chemistry.chemical_compound Cricetulus Report 010608 biotechnology Animals Immunology and Allergy Molecule Disulfides Staphylococcal Protein A Hydrogen peroxide Aqueous solution biology Chemistry Disulfide bond reduction Antibodies Monoclonal Hydrogen Peroxide Combinatorial chemistry Recombinant Proteins 030104 developmental biology Immunoglobulin G biology.protein Aeration Antibody Oxidation-Reduction |
| Zdroj: | mAbs. 10:500-510 |
| ISSN: | 1942-0870 1942-0862 |
| DOI: | 10.1080/19420862.2018.1424609 |
| Popis: | During large-scale monoclonal antibody manufacturing, disulfide bond reduction of antibodies, which results in generation of low molecule weight species, is occasionally observed. When this happens, the drug substance does not meet specifications. Many investigations have been conducted across the biopharmaceutical industry to identify the root causes, and multiple strategies have been proposed to mitigate the problem. The reduction is correlated with the release of cellular reducing components and depletion of dissolved oxygen before, during, and after harvest. Consequently, these factors can lead to disulfide reduction over long-duration storage at room temperature prior to Protein A chromatography. Several strategies have been developed to minimize antibody reduction, including chemical inhibition of reducing components, maintaining aeration before and after harvest, and chilling clarified harvest during holding. Here, we explore the use of hydrogen peroxide in clarified harvest bulk or cell culture fluid as a strategy to prevent disulfide reduction. A lab-scale study was performed to demonstrate the effectiveness of hydrogen peroxide in preventing antibody reduction using multiple IgG molecules. Studies were done to define the optimal concentration of hydrogen peroxide needed to avoid unnecessary oxidization of the antibody products. We show that adding a controlled amount of hydrogen peroxide does not change product quality attributes of the protein. Since hydrogen peroxide is soluble in aqueous solutions and decomposes into water and oxygen, there is no additional burden involved in removing it during the downstream purification steps. Due to its ease of use and minimal product impact, we demonstrate that hydrogen peroxide treatment is a powerful, simple tool to quench reducing potential by simply mixing it with harvested cell culture fluid. |
| Databáze: | OpenAIRE |
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