Dilara Baş¹, Emre Burak Erkal¹, Ali Özhan Aytekin², Deniz Demirhan^1,3*

¹Biotechnology Group, Turgut Pharmaceuticals, İstanbul, Turkey
²Genetics and Bioengineering Department, Faculty of Engineering, Yeditepe University, Atasehir, İstanbul, Turkey
³Department of Natural Sciences, Faculty of Engineering, Acibadem Mehmet Ali Aydinlar University, Istanbul, Turkey

*Corresponding Author: Deniz Demirhan, Turgut Pharmaceuticals Biotechnology Group/Department of Natural Sciences, Faculty of Engineering, Acibadem Mehmet Ali Aydınlar University, İstanbul, Turkey.

Received:July 28, 2021 Published: September 24, 2021

Citation: Deniz Demirhan., et al. “Indirect (Mirror) Laryngoscopy: Valuable Laryngological Skill Going into Extinction?". Acta Scientific Medical Sciences 5.10 (2021): 144-155.

Abstract

  Therapeutic monoclonal antibodies (mAbs) are target-specific proteins used for patients especially suffering from cancer, autoimmune, and inflammatory diseases. Due to the success rate of the mAbs, they are the fastest-growing sector in the biopharmaceutical industry. To be used for the therapeutic purposes, the monoclonal antibodies have to be high purity. A variety of different downstream technologies have recently been evolved to provide high quality monoclonal antibody and biosimilar development with low cost in a shorter time. Recent advancements of high-throughput process (HTP) technology are employed for process development for saving time, investment, and cost. HTP allows running large number of experiments in a minimal scale. Incorporation of HTP technology with Design of Experiment (DoE) is systematic integration for effective and rapid screening of optimum experimental conditions.

  Development of purification process for mAbs requires the consideration of many different parameters including removal of impurities and ready availability of raw materials for large-scale production. In this study, a downstream process was developed for one of the best-selling biosimilar mAb (TUR02) coupling HTP technology with DoE tools. TUR02 is an IgG1 which has a high tendency to aggregate. In order to develop protein A chromatography, firstly appropriate elution buffer had been selected using micro-volume columns. Large range of parameters were screened as buffer type (acetic acid, citric acid, sodium acetate), the concentration of buffer (10 mM-150 mM), and three different Protein A resins. Citric acid (10mM) at pH 3.5 was chosen as suitable elution buffer. A resin with base matrix of methacrylic polymer was chosen from the HTP study and a lab scale experiment with 1 ml column was done to prove the success of the scalability of the HTP technology. After protein A chromatography, HTP technology was applied to cation exchange chromatography to screen different conditions to be able to further decrease the impurity level. Flow through with overloading mode was followed with the cation exchange screening. Concentration of sodium phosphate-based buffer (10 mM – 50 mM), working pH and comparison of strong and weak cation exchange resin were screened by coupling HTP technology and DoE tools. Strong cation exchange resin, 38 mM sodium phosphate and pH 4.5 were selected due to its impurity reduction strength. Scalability of the process was successfully proven using 1 ml pre-packed column. With these conditions, the aggregate and HCP levels were 0.52 and 612 and 0.32 and 234 for 0.1 ml and 1 ml columns, respectively.

Keywords: Monoclonal Antibodies; Chromatography; High-Throughput Technology; Design of Experiment; Protein A Ligand; Cation Exchange

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Copyright: © 2021 Deniz Demirhan., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.