Explore the Journey of Therapeutic Antibody Production

The therapeutic antibody journey starts with a specific antibody gene sequence, which is optimized and inserted into the genome of host cells – typically Chinese hamster ovary (CHO) cells. This development process typically takes 6–12 months and involves numerous analytical tests – including sequencing and binding assays – to ensure the cells produce antibodies with the correct sequence, structure and functional properties. Therapeutic antibodies represent one of the most important advances of the modern biopharmaceutic landscape, with over 100 approved treatments and thousands more in development.1 These complex proteins can precisely target disease mechanisms while minimizing side effects, making them ideal therapeutic tools against hard-to-treat diseases. This infographic explores the journey involved in therapeutic antibody production – from initial engineering to final purification. Engineered cells then undergo extensive screening and characterization, with only one candidate making it through to become a production cell line out of thousands of possibilities.2 The selected cells must demonstrate key criteria, including: Consistent growth Genetic stability High productivity Engineering the perfect producer THERAPEUTIC ANTIBODY The Development Journey of a Initial cell culture and expansion Harvesting the desired product Purification and polishing Final processing This phase begins in small shake flasks containing just a few hundred milliliters of nutrient media. These volumes are gradually increased and, depending on manufacturing requirements, can be carried out in production vessels holding 10,000–20,000 liters.3 Throughout the scale-up process, various parameters are monitored to maintain optimal conditions for cell growth and antibody production. A combination of automated and manual technology is used to continuously monitor and adjust pH, temperature, dissolved oxygen and nutrient levels. This culture phase typically runs for 10–14 days, during which cells multiply and produce antibodies that accumulate in the culture medium. After primary recovery, depth filtration steps remove finer particles. Throughout this process, conditions are carefully controlled to prevent antibody degradation or aggregation, maintaining the temperature, pH and ionic conditions within narrow ranges. Primary recovery operations remove intact cells and larger debris, typically using industrial centrifuges or specialized filtration systems. The entire process operates under aseptic conditions to prevent contamination, with extensive in-process testing to verify product quality. The harvested fluid is a complex mixture containing the target antibody along with host cell proteins, DNA and media components. It must meet strict quality specifications before advancing to purification. This stage begins with capture chromatography, typically using Protein A resins – specialized materials that selectively bind to the constant (Fc) region of antibodies. This method can achieve over 95% purity in a single step, concentrating the antibody while removing the majority of contaminants. This phase begins with viral filtration – specialized nanometerscale filters that provide an additional safety measure by removing potential viral contaminants. The antibody solution then undergoes ultrafiltration and diafiltration to concentrate the antibodies. The formulated drug substance is then sterile filtered and aseptically added to vials or pre-filled syringes using high-precision automated equipment. When sufficient volumes of antibodies have accumulated and reached peak concentration, the harvesting process begins. The purification phase transforms a complex biological mixture into a highly pure product through a series of chromatography steps. Each step is designed to separate the target antibody from process-related impurities. Ion exchange Hydrophobic interaction Size exclusion Production process STAGE 1 STAGE 3 Purity >99% Purity Sample Target antibodies Protein A resins 95% purity WASTE LARGE DEBRIS Following capture, intermediate and polishing chromatography steps employ different separation principles to remove remaining impurities. The entire purification process is validated to consistently deliver a final product meeting stringent purity requirements: typically > 99% pure, with impurities measured in parts per million. Formulation is then used to combine the antibody with carefully selected excipients –such as buffering agents, stabilizers and tonicity modifiers – to maintain stability, prevent aggregation and ensure patient comfort during administration. References 1. Lyu X, Zhao Q, Hui J, et al. The global landscape of approved antibody therapies. Antib Ther. 2022;5(4):233–257. doi: 10.1093/abt/tbac021 2. B ailly M, Mieczkowski C, Juan V, et al. Predicting antibody developability profiles through early stage discovery screening. mAbs. 2020;12(1):1743053. doi: 10.1080/19420862.2020.1743053 3. Kelley B. Industrialization of mAb production technology: The bioprocessing industry at a crossroads. mAbs. 2009;1(5):443–452. doi: https://doi.org/10.4161/mabs.1.5.9448 The entire process, from initial cell culture to final product, operates under stringent good manufacturing practice (GMP) regulations to ensure consistent quality and patient safety. Each batch undergoes extensive quality control testing including identity, purity, potency, sterility and endotoxin testing, before being labeled, packaged and released for distribution. 1 2 3 4 5 6 8 7 9 10 11 12 13 14 15 16 17 18 Steven Gibney, PhD Illustrated by Alisha Vroom